Purine inhibitors of fructose 1,6-bisphosphatase

ABSTRACT

Novel purine compounds of the following structure and their use as fructose-1,6-bisphosphatase inhibitors is described.                    
     wherein 
     A is selected from the group consisting of —NR 8   2 , —NHSO 2 R 3 , —OR 5 , —SR 5 , halo, lower alkyl, —CON(R 4 ) 2 , guanidino, amidino, —H, and perhaloalkyl; 
     E is selected from the group consisting of —H, halo, lower alkylthio, lower perhaloalkyl, lower alkyl, lower alkenyl, lower alkynyl, lower alkoxy, —CN, and —NR 7   2 ; 
     X is selected from the group consisting of -alk-NR—, alkylene, alkenylene, alkynylene, arylene, heteroarylene, -alk-NR-alk-, -alk-O-alk-, -alk-S-alk-, -alk-S—, alicyclicene, heteroalicyclicene,  1,1 -dihaloalkylene, —C(O)-alk-, —NR—C(O)—NR′—, -alk-NR—C(O)—, -alk-C(O)—NR—, —Ar-alk-, and -alk-Ar—, all optionally substituted, wherein each R and R′ is independently selected from —H and lower alkyl, and wherein each “alk” and “Ar” is an independently selected alkylene or arylene, respectively; 
     Y is selected from the group consisting of —H, alkyl, alkenyl, alkynyl, aryl, alicyclic, heteroalicyclic, aralkyl, aryloxyalkyl, alkoxyalkyl, —C(O)R 3 , —S(O) 2 R 3 , —C(O)—OR 3 , —CONHR 3 , —NR 2   2 , and —OR 3 , all except H are optionally substituted; and 
     pharmaceutically acceptable prodrugs and salts thereof.

RELATED APPLICATIONS

This application claims the benefit of and is a continuation-in-part ofU.S. Provisional Application No. 60/040,623, filed Mar. 7, 1997, nowabandoned.

FIELD OF THE INVENTION

This invention relates to novel purine compounds that are inhibitors ofFructose-1,6-bisphosphatase at the AMP site. The invention also relatesto the preparation and use of these purine analogs in the treatment ofdiabetes, and other diseases where the inhibition of gluconeogenesis,control of blood glucose levels, reduction in glycogen stores, orreduction in insulin levels is beneficial.

BACKGROUND AND INTRODUCTION TO THE INVENTION

Diabetes mellitus (or diabetes) is one of the most prevalent diseases inthe world today. Diabetes patients have been divided into two classes,namely type I or insulin-dependent diabetes mellitus and type II ornon-insulin dependent diabetes mellitus (NIDDM). Non-insulin-dependentdiabetes mellitus (NIDDM) accounts for approximately 90% of alldiabetics and is estimated to affect 12-14 million adults in the U.S.alone (6.6% of the population). NIDDM is characterized by both fastinghyperglycemia and exaggerated postprandial increases in plasma glucoselevels. NIDDM is associated with a variety of long-term complications,including microvascular diseases such as retinopathy, nephropathy andneuropathy, and macrovascular diseases such as coronary heart disease.Numerous studies in animal models demonstrate a causal relationshipbetween long term complications and hyperglycemia. Recent results fromthe Diabetes Control and Complications Trial (DCCT) and the StockholmProspective Study demonstrate this relationship for the first time inman by showing that insulin-dependent diabetics with tighter glycemiccontrol are at substantially lower risk for development and progressionof these complications. Tighter control is also expected to benefitNIDDM patients.

Current therapies used to treat NIDDM patients entail both controllinglifestyle risk factors and pharmaceutical intervention. First-linetherapy for NIDDM is typically a tightly-controlled regimen of diet andexercise since an overwhelming number of NIDDM patients are overweightor obese (≈67%) and since weight loss can improve insulin secretion,insulin sensitivity and lead to normoglycemia. Normalization of bloodglucose occurs in less than 30% of these patients due to poor complianceand poor response. Patients with hyperglycemia not controlled by dietalone are subsequently treated with oral hypoglycemics or insulin. Untilrecently, the sulfonylureas were the only class of oral hypoglycemicagents available for NIDDM. Treatment with sulfonylureas leads toeffective blood glucose lowering in only 70% of patients and only 40%after 10 years of therapy. Patients that fail to respond to diet andsulfonylureas are subsequently treated with daily insulin injections togain adequate glycemic control.

Although the sulfonylureas represent a major therapy for NIDDM patients,four factors limit their overall success. First, as mentioned above, alarge segment of the NIDDM population do not respond adequately tosulfonylurea therapy (i.e. primary failures) or become resistant (i.e.secondary failures). This is particularly true in NIDDM patients withadvanced NIDDM since these patients have severely impaired insulinsecretion. Second, sulfonylurea therapy is associated with an increasedrisk of severe hypoglycemic episodes. Third, chronic hyperinsulinemiahas been associated with increased cardiovascular disease although thisrelationship is considered controversial and unproven. Last,sulfonylureas are associated with weight gain, which leads to worseningof peripheral insulin sensitivity and thereby can accelerate theprogression of the disease.

Recent results from the U.K. Diabetes prospective study also showed thatpatients undergoing maximal therapy of a sulfonylurea, metformin, or acombination of the two, were unable to maintain normal fasting glycemiaover the six year period of the study. U.K. Prospective Diabetes Study16. Diabetes, 44:1249-158 (1995). These results further illustrate thegreat need for alternative therapies. Three therapeutic strategies thatcould provide additional health benefits to NIDDM patients beyond thecurrently available therapies, include drugs that would: (i) prevent theonset of NIDDM; (ii) prevent diabetic complications by blockingdetrimental events precipitated by chronic hyperglycemia; or (iii)normalize glucose levels or at least decrease glucose levels below thethreshold reported for microvascular and macrovascular diseases.

Hyperglycemia in NIDDM is associated with two biochemical abnormalities,namely insulin resistance and impaired insulin secretion. The relativeroles of these metabolic abnormalities in the pathogenesis of NIDDM hasbeen the subject of numerous studies over the past several decades.Studies of offspring and siblings of NIDDM patients, mono- and dizygotictwins, and ethnic populations with high incidence of NIDDM (e.g. PimaIndians) strongly support the inheritable nature of the disease.

Despite the presence of insulin resistance and impaired insulinsecretion, fasting blood glucose (FBG) levels remain normal inpre-diabetic patients due to a state of compensatory hyperinsulinemia.Eventually, however, insulin secretion is inadequate and fastinghyperglycemia ensues. With time insulin levels decline. Progression ofthe disease is characterized by increasing FBG levels and declininginsulin levels.

Numerous clinical studies have attempted to define the primary defectthat accounts for the progressive increase in FBG. Results from thesestudies indicate that excessive hepatic glucose output (HGO) is theprimary reason for the elevation in FBG with a significant correlationfound for HGO and FBG once FBG exceeds 140 mg/dL. Kolterman, et al., J.Clin. Invest. 68:957, (1981); DeFronzo Diabetes 37:667 (1988).

HGO comprises glucose derived from breakdown of hepatic glycogen(glycogenolysis) and glucose synthesized from 3-carbon precursors(gluconeogenesis). A number of radioisotope studies and several studiesusing ¹³C-NMR spectroscopy have shown that gluconeogenesis contributesbetween 50-100% of the glucose produced by the liver in thepostabsorptive state and that gluconeogenesis flux is excessive (2- to3-fold) in NIDDM patients. Magnusson, et al. J. Clin. Invest.90:1323-1327 (1992); Rothman, et al., Science 254: 573-76 (1991);Consoli, et al. Diabetes 38:550-557 (1989).

Gluconeogenesis from pyruvate is a highly regulated biosynthetic pathwayrequiring eleven enzymes (FIG. 1). Seven enzymes catalyze reversiblereactions and are common to both gluconeogenesis and glycolysis. Fourenzymes catalyze reactions unique to gluconeogenesis, namely pyruvatecarboxylase, phosphoenolpyruvate carboxykinase,fructose-1,6-bisphosphatase and glucose-6-phosphatase. Overall fluxthrough the pathway is controlled by the specific activities of theseenzymes, the enzymes that catalyzed the corresponding steps in theglycolytic direction, and by substrate availability. Dietary factors(glucose, fat) and hormones (insulin, glucagon, glucocorticoids,epinephrine) coordinatively regulate enzyme activities in thegluconeogenesis and glycolysis pathways through gene expression andpost-translational mechanisms.

Of the four enzymes specific to gluconeogenesis,fructose-1,6-bisphosphatase (hereinafter “FBPase”) is the most suitabletarget for a gluconeogenesis inhibitor based on efficacy and safetyconsiderations. Studies indicate that nature uses the FBPase/PFK cycleas a major control point (metabolic switch) responsible for determiningwhether metabolic flux proceeds in the direction of glycolysis orgluconeogenesis. Claus, et al., Mechanisms of Insulin Action, Belfrage,P. editor, pp.305-321, Elsevier Science 1992; Regen, et al. J. Theor.Biol., 111:635-658 (1984); Pilkis, et al. Annu. Rev. Biochem, 57:755-783(1988). FBPase is inhibited by fructose-2,6-bisphosphate in the cell.Fructose-2,6-bisphosphate binds to the substrate site of the enzyme. AMPbinds to an allosteric site on the enzyme.

Synthetic inhibitors of FBPase have also been reported. McNiel reportedthat fructose-2,6-bisphosphate analogs inhibit FBPase by binding to thesubstrate site. J. Am. Chem. Soc. 106:7851 (1984); U.S. Pat. No.4,968,790 (1984). These compounds, however, were relatively weak and didnot inhibit glucose production in hepatocytes presumably due to poorcell penetration.

Gruber reported that some nucleosides can lower blood glucose in thewhole animal through inhibition of FBPase. These compounds exert theiractivity by first undergoing phosphorylation to the correspondingmonophosphate. EP 0 427 799 B1.

Gruber et al. U.S. Pat. No. 5,658,889 described the use of inhibitors ofthe AMP site of FBPase to treat diabetes.

European patent application EP 0 632 048 A1 discloses certain ethylphosphonates of purine derivatives for use as antiviral andantineoplastic agents. These structures differ from the claimedcompounds because they have no substitution on the C-8 of the purine.There is no suggestion that these compounds are inhibitors of FBPase.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a scheme depicting the eleven enzymes of the gluconeogenesispathway.

FIG. 2 shows the dose-dependent inhibition of hPFBPase AMP and compounds2.7, 2.13, and 2.5.

FIG. 3 shows the displacement of ³H-AMP from hPFBPase by ZMP andcompound 2.2.

FIG. 4A depicts the reduction of glucose production fromdihydroxyacetone in rat hepatocytes treated with compound 2.7.

FIG. 4B depicts the increase in the amount of fructose-1,6-bisphosphatein rat hepatocytes exposed to dihydroxyacetone and treated with compound2.7.

FIG. 5 depicts a dose-dependent inhibition of glucose production in rathepatocytes exposed to lactate and pyruvate by compounds 2.7 and 2.1.

FIG. 6 shows the effect of various doses of compound 16.4 on bloodglucose levels in 18 hour fasted, normal rats.

FIG. 7 is a bar graph depicting the reduction in blood glucose levels in18 hour fasted normal rats treated with compound 2.7 given at a dose of20 mg/kg i.p.

FIG. 8 is a bar graph depicting the increased accumulation offructose-1,6-bisphosphate in the liver of 18-hour fasted rats treatedwith compound 2.7.

FIG. 9A is a bar graph depicting a reduction in blood glucose levels in24 hour fasted Zucker Diabetic Fatty rats treated with compound 2.7.

FIG. 9B is a bar graph depicting the percentage change in blood glucoselevels in 24 hour fasted Zucker Diabetic Fatty rats treated withcompound 2.7.

FIG. 10 depicts the inhibition of gluconeogenesis from ¹⁴C-bicarbonatein 24 hour fasted Diabetic Fatty rats treated with compound 2.7.

FIG. 11A depicts the dose-dependent inhibition of glucose production inrat hepatocytes by compound 16.4, a prodrug of compound 2.7.

FIG. 11B shows the intracellular generation of compound 2.7 in rathepatocytes treated with compound 16.4, a prodrug, to inhibit glucoseproduction in rat hepatocytes.

SUMMARY OF THE INVENTION

The present invention is directed towards novel purine compounds whichbind the AMP site and are potent FBPase inhibitors. In another aspect,the present invention is directed to the preparation of these novelpurine compounds and to the in vitro and in vivo FBPase inhibitoryactivity of these compounds. Another aspect of the present invention isdirected to the clinical use of the novel FBPase inhibitors as a methodof treatment or prevention of diseases responsive to inhibition ofgluconeogenesis and in diseases responsive to lowered blood glucoselevels.

Gruber et al. U.S. patent application Ser. No. 08/355,836, now issuedU.S. Pat. No 5,658,889, described the use of inhibitors of the AMP siteof FBPase to treat diabetes.

The compounds are also useful in treating or preventing excess glycogenstorage diseases and insulin dependent diseases such as cardiovasculardiseases including atherosclerosis.

The invention comprises the novel purine analogs as specified below informula 1. Also included in the scope of the present invention areprodrugs of the compounds of formula 1.

Since these compounds may have asymmetric centers, the present inventionis directed not only to racemic mixtures of these compounds, but also toindividual stereoisomers. The present invention also includespharmaceutically acceptable and/or useful salts of the compounds offormula 1, including acid addition salts and basic salts. The presentinventions also encompass prodrugs of compounds of formula 1.

Definitions

In accordance with the present invention and as used herein, thefollowing terms are defined with the following meanings, unlessexplicitly stated otherwise.

The term “aryl” refers to aromatic groups which have at least one ringhaving a conjugated pi electron system and includes carbocyclic aryl,heterocyclic aryl and biaryl groups, all of which may be optionallysubstituted.

Carbocyclic aryl groups are groups wherein the ring atoms on thearomatic ring are carbon atoms. Carbocyclic aryl groups includemonocyclic carbocyclic aryl groups and polycyclic or fused compoundssuch as optionally substituted naphthyl groups.

Heterocyclic aryl groups are groups having from 1 to 4 heteroatoms asring atoms in the aromatic ring and the remainder of the ring atomsbeing carbon atoms. Suitable heteroatoms include oxygen, sulfur, andnitrogen. Suitable heteroaryl groups include furanyl, thienyl, pyridyl,pyrrolyl, N-lower alkyl pyrrolyl, pyridyl-N-oxide, pyrimidyl, pyrazinyl,imidazolyl, and the like, all optionally substituted.

The term “biaryl” represents aryl groups containing more than onearomatic ring including both fused ring systems and aryl groupssubstituted with other aryl groups.

The term “alicyclic” means compounds which combine the properties ofaliphatic and cyclic compounds and include but are not limited toaromatic, cycloalkyl and bridged cycloalkyl compounds. The cycliccompound includes heterocycles. Cyclohexenylethyl, cyclohexanylethyl,and norbornyl are suitable alicyclic groups. Such groups may beoptionally substituted.

The term “optionally substituted” or “substituted” includes groupssubstituted by one to four substituents, independently selected fromlower alkyl, lower aryl, lower aralkyl, lower alicyclic, hydroxy, loweralkoxy, lower aryloxy, perhaloalkoxy, aralkoxy, heteroaryl,heteroaryloxy, heteroarylalkyl, heteroaralkoxy, azido, amino, guanidino,halogen, lower alkylthio, oxo, ketone, carboxy esters, carboxyl,carboxamido, nitro, acyloxy, alkylamino, aminoalkyl, alkylaminoaryl,alkylaryl, alkylaminoalkyl, alkoxyaryl, arylamino, aralkylamino,phosphonate, sulfonate, carboxamidoalkylaryl, carboxamidoaryl,hydroxyalkyl, haloalkyl, alkylaminoalkylcarboxy, aminocarboxamidoalkyl,cyano, lower alkoxyalkyl, and lower perhaloalkyl.

The term “aralkyl” refers to an alkyl group substituted with an arylgroup. Suitable aralkyl groups include benzyl, picolyl, and the like,and may be optionally substituted.

The term “lower” referred to herein in connection with organic radicalsor compounds respectively defines such as with up to and including 10,preferably up to and including 6, and advantageously one to four carbonatoms. Such groups may be straight chain, branched, or cyclic.

The terms “arylamino” (a), and “aralkylamino” (b), respectively, referto the group —NRR′ wherein respectively, (a) R is aryl and R′ ishydrogen, alkyl, aralkyl or aryl, and (b) R is aralkyl and R′ ishydrogen or aralkyl, aryl, alkyl.

The term “acyl” refers to —C(O)R where R is alkyl and aryl.

The term “carboxy esters” refers to —C(O)OR where R is alkyl, aryl,aralkyl, and alicyclic, all optionally substituted.

The term “oxa” refers to ═O in an alkyl group.

The term “alkylamino” refers to —NRR′ where R and R′ are independentlyselected from hydrogen or alkyl.

The term “carbonylamine” or “carbonylamino” refers to —CONR₂ where eachR is independently hydrogen or alkyl.

The term “halogen” or “halo” refers to —F, —Cl, —Br and —I.

The term “oxyalkylamino” refers to —O-alk-NR—, where “alk” is analkylene group and R is H or alkyl.

The term “alkylaminoalkylcarboxy” refers to the group-alk-NR-alk-C(O)—O— where “alk” is an alkylene group, and R is a H orlower alkyl.

The term “alkylaminocarbonyl” refers to the group -alk-NR—C(O)— where“alk” is an alkylene group, and R is a H or lower alkyl.

The term “oxyalkyl” refers to the group —O-alk- where “alk” is analkylene group.

The term “alkylcarboxyalkyl” refers to the group -alk-C(O)—O-alkyl whereeach alk is independently an alkylene group.

The term “alkyl” refers to saturated aliphatic groups includingstraight-chain, branched chain and cyclic groups. Alkyl groups may beoptionally substituted.

The term “bidentate” refers to an alkyl group that is attached by itsterminal ends to the same atom to form a cyclic group. For example,propylene imine contains a bidentate propylene group.

The term “cyclic alkyl” refers to alkyl groups that are cyclic.

The term “heterocyclic” and “heterocyclic alkyl” refer to cyclic alkylgroups containing at least one heteroatom. Suitable heteroatoms includeoxygen, sulfur, and nitrogen. Heterocyclic groups may be attachedthrough a heteroatom or through a carbon atom in the ring.

The term “alkenyl” refers to unsaturated groups which contain at leastone carbon-carbon double bond and includes straight-chain,branched-chain and cyclic groups. Alkene groups may be optionallysubstituted.

The term “alkynyl” refers to unsaturated groups which contain at leastone carbon-carbon triple bond and includes straight-chain,branched-chain and cyclic groups. Alkyne groups may be optionallysubstituted.

The term “alkylene” refers to a divalent straight chain, branched chainor cyclic saturated aliphatic radical.

The term “acyloxy” refers to the ester group —O—C(O)R, where R is H,alkyl, alkenyl, alkynyl, aryl, aralkyl, or alicyclic.

The term “alkylaryl” refers to the group -alk-aryl- where “alk” is analkylene group. “Lower alkylaryl” refers to such groups where alkyleneis lower alkyl.

The term “alkylamino” refers to the group -alk-NR— wherein “alk” is analkylene group.

The term “alkyl(carboxyl)” refers to carboxyl substituted off the alkylchain. Similarly, “alkyl(hydroxy)”, “alkyl(phosphonate)”, and“alkyl(sulfonate)” refers to substituents off the alkyl chain.

The term “alkylaminoalkyl” refers to the group -alk-NR-alk- wherein each“alk” is an independently selected alkylene, and R is H or lower alkyl.“Lower alkylaminoalkyl” refers to groups where each alkylene group islower alkyl.

The term “alkylaminoaryl” refers to the group -alk-NR-aryl- wherein“alk”is an alkylene group. In “lower alkylaminoaryl”, the alkylene groupis lower alkyl.

The term “alkyloxyaryl” refers to an alkylene group substituted with anaryloxy group. In “lower alkyloxyaryl”, the alkylene group is loweralkyl.

The term “alkylacylamino” refers to the group -alk-N—(COR)— wherein alkis alkylene and R is lower alkyl. In “lower alkylacylamino”, thealkylene group is lower alkyl.

The term “alkoxyalkylaryl” refers to the group -alk-O-alk-aryl- whereineach “alk” is independently an alkylene group. “Lower alkoxalkylaryl”refers to such groups where the alkylene group is lower alkyl.

The term “alkylacylaminoalkyl” refers to the group -alk-N—(COR)-alk-where each alk is an independently selected alkylene group. In “loweralkylacylaminoalkyl” the alkylene groups are lower alkyl.

The term “alkoxy” refers to the group -alk-O— wherein alk is an alkylenegroup.

The term “alkoxyalkyl” refers to the group -alk-O-alk- wherein each alkis an independently selected alkylene group. In “lower alkoxyalkyl”,each alkylene is lower alkyl.

The term “alkylthio” refers to the group -alk-S— wherein alk is alkylenegroup.

The term “alkylthioalkyl” refers to the group -alk-S-alk- wherein eachalk is an independently selected alkylene group. In “loweralkylthioalkyl” each alkylene is lower alkylene.

The term “aralkylamino” refers to an amine substituted with an aralkylgroup.

The term “alkylcarboxamido” refers to the group -alk-C(O)N(R)— whereinalk is an alkylene group and R is H or lower alkyl.

The term “alkylcarboxamidoalkyl” refers to the group -alk-C(O)N(R)-alk-wherein each alk is an independently selected alkylene group and R islower alkyl. In “lower alkylcarboxamidoalkyl” each alkylene is loweralkyl.

The term “alkylcarboxamidoalkylaryl” refers to the group-alk₁-C(O)—NH-alk₂Ar— wherein alk₁ and alk₂ are independently selectedalkylene groups and alk₂ is substituted with an aryl group, Ar. In“lower alkylcarboxamidoalkylaryl”, each alkylene is lower alkyl.

The term “hteteroalicyclic” refers to an alicyclic group having 1 to 4heteroatoms in the ring selected from nitrogen, sulfur, phosphorus andoxygen.

The term “aminocarboxamidoalkyl” refers to the group —NH—C(O)—N(R)—Rwherein each R is an independently selected alkyl group. “Loweraminocarboxamidoalkyl” refers to such groups wherein each R is loweralkyl.

The term “heteroarylalkyl” refers to an alkyl group substituted with aheteroaryl group.

The term “perhalo” refers to groups wherein every C—H bond has beenreplaced with a C-halo bond on an aliphatic or aryl group. Suitableperhaloalkyl groups include —CF₃ and —CFCl₂.

The term “guanidino” refers to both —NR—C(NR)—NR₂ as well as —N═C(NR₂)₂where each R group is independently selected from the group of —H,alkyl, alkenyl, alkynyl, aryl, and alicyclic, all optionallysubstituted.

The term “amidino” refers to —C(NR)—NR₂ where each R group isindependently selected from the group of —H, alkyl, alkenyl, alkynyl,aryl, and alicyclic, all optionally substituted.

The term “pharmaceutically acceptable salt” includes salts of compoundsof formula 1 and its prodrugs derived from the combination of a compoundof this invention and an organic or inorganic acid or base.

The term “prodrug” as used herein refers to any compound that whenadministered to a biological system generates the “drug” substanceeither as a result of spontaneous chemical reaction(s) or by enzymecatalyzed or metabolic reaction(s). Reference is made to variousprodrugs such as acyl esters, carbonates, and carbamates, includedherein. The groups illustrated are exemplary, not exhaustive, and oneskilled in the art could prepare other known varieties of prodrugs. Suchprodrugs of the compounds of formula 1, fall within the scope of thepresent invention.

The term “prodrug ester” as employed herein includes, but is not limitedto, the following groups and combinations of these groups:

[1] Acyloxyalkyl esters which are well described in the literature(Farquhar et al., J. Pharm. Sci. 72, 324-325 (1983)) and are representedby formula A

wherein

R, R′, and R″ are independently H, alkyl, aryl, alkylaryl, andalicyclic; (see WO 90/08155; WO 90/10636).

[2] Other acyloxyalkyl esters are possible in which an alicyclic ring isformed such as shown in formula B. These esters have been shown togenerate phosphorus-containing nucleotides inside cells through apostulated sequence of reactions beginning with deesterification andfollowed by a series of elimination reactions (e.g. Freed et al.,Biochem. Pharm. 38: 3193-3198 (1989)).

wherein

R is —H, alkyl, aryl, alkylaryl, alkoxy, aryloxy, alkylthio, arylthio,alkylamino, arylamino, cycloalkyl, or alicyclic.

[3] Another class of these double esters known asalkyloxycarbonyloxymethyl esters, as shown in formula A, where R isalkoxy, aryloxy, alkylthio, arylthio, alkylamino, and arylamino; R′, andR″ are independently H, alkyl, aryl, alkylaryl, and alicyclic, have beenstudied in the area of β-lactam antibiotics (Tatsuo Nishimura et al. J.Antibiotics, 1987, 40(1), 81-90; for a review see Ferres, H., Drugs ofToday, 1983,19, 499.). More recently Cathy, M. S., et al. (Abstract fromAAPS Western Regional Meeting, April, 1997) showed that thesealkyloxycarbonyloxymethyl ester prodrugs on(9-[(R)-2-phosphonomethoxy)propyl]adenine (PMPA) are bioavailable up to30% in dogs.

[4] Aryl esters have also been used as phosphonate prodrugs (e.g. Erion,DeLambert et al., J. Med. Chem. 37: 498, 1994; Serafinowska et al., J.Med. Chem. 38: 1372, 1995). Phenyl as well as mono and poly-substitutedphenyl phosphonate ester prodrugs have generated the parent phosphonicacid in studies conducted in animals and in man (Formula C). Anotherapproach has been described where Y is a carboxylic ester ortho to thephosphate. Khamnei and Torrence, J. Med. Chem.; 39:4109-4115 (1996).

wherein

Y is H, alkyl, aryl, alkylaryl, alkoxy, acetoxy, halogen, amino,alkoxycarbonyl, hydroxy, cyano, alkylamino, and alicyclic.

[5] Benzyl esters have also been reported to generate the parentphosphonic acid. In some cases, using substituents at the para-positioncan accelerate the hydrolysis. Benzyl analogs with 4-acyloxy or4-alkyloxy group [Formula D, X═H, OR or O(CO)R or O(CO)OR] can generatethe 4-hydroxy compound more readly through the action of enzymes, e.g.oxidases, esterases, etc. Examples of this class of prodrugs aredescribed by Mitchell et al., J. Chem. Soc. Perkin Trans. I 2345 (1992);Brook, et al. WO 91/19721.

wherein

X and Y are independently H, alkyl, aryl, alkylaryl, alkoxy, acetoxy,hydroxy, cyano, nitro, perhaloalkyl, halo, or alkyloxycarbonyl; and

R′ and R″ are independently H, alkyl, aryl, alkylaryl, halogen, andalicyclic.

[6] Thio-containing phosphonate ester prodrugs have been described thatare useful in the delivery of FBPase inhibitors to hepatocytes. Thesephosphonate ester prodrugs contain a protected thioethyl moiety as shownin formula E. One or more of the oxygens of the phosphonate can beesterified. Since the mechanism that results in de-esterificationrequires the generation of a free thiolate, a variety of thiolprotecting groups are possible. For example, the disulfide is reduced bya reductase-mediated process (Puech et al., Antiviral Res., 22: 155-174(1993)). Thioesters will also generate free thiolates afteresterase-mediated hydrolysis. Benzaria, et al., J. Med. Chem., 39:4958(1996). Cyclic analogs are also possible and were shown to liberatephosphonate in isolated rat hepatocytes. The cyclic disulfide shownbelow has not been previously described and is novel.

wherein

Z is alkylcarbonyl, alkoxycarbonyl, arylcarbonyl, aryloxycarbonyl, oralkylthio.

Other examples of suitable prodrugs include proester classes exemplifiedby Biller and Magnin (U.S. Pat. No. 5,157,027); Serafinowska et al. (J.Med. Chem. 38,1372 (1995)); Starrett et al. (J. Med. Chem. 37,1857(1994)); Martin et al. J. Pharm. Sci. 76,180 (1987); Alexander et al.,Collect. Czech. Chem. Commun, 59, 1853 (1994)); and EPO patentapplication 0 632 048 A1. Some of the structural classes described areoptionally substituted, including fused lactones attached at the omegaposition and optionally substituted 2-oxo-1,3-dioxolenes attachedthrough a methylene to the phosphorus oxygen such as:

wherein

R is —H, alkyl, cycloalkyl, or alicyclic; and

wherein Y is —H, alkyl, aryl, alkylaryl, cyano, alkoxy, acetoxy,halogen, amino, alkylamino, alicyclic, and alkoxycarbonyl.

[7] Propyl phosphonate ester prodrugs can also be used to deliver FBPaseinhibitors into hepatocytes. These phosphonate ester prodrugs maycontain a hydroxyl and hydroxyl group derivatives at the 3-position ofthe propyl group as shown in formula F. The R and X groups can form acyclic ring system as shown in formula F. One or more of the oxygens ofthe phosphonate can be esterified.

wherein

R is alkyl, aryl, heteroaryl;

X is hydrogen, alkylcarbonyloxy, alkyloxycarbonyloxy; and

Y is alkyl, aryl, heteroaryl, alkoxy, alkylamino, alkylthio, halogen,

hydrogen, hydroxy, acetoxy, amino.

[8] The cyclic propyl phosphonate esters as in Formula G are shown toactivate to phosphonic acids. The activation of prodrug can bemechanistically explained by in vivo oxidation and elimination steps.These prodrugs inhibit glucose production in isolated rat hepatocytesand are also shown to deliver FBPase inhibitors to the liver followingoral administration.

wherein

V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or

together V and Z are connected to form a cyclic group containing 3-5atoms, optionally 1 heteroatom, substituted with hydroxy, acyloxy,alkoxycarboxy, or aryloxycarboxy attached to a carbon atom that is threeatoms from an oxygen attached to the phosphorus; or

together V and W are connected to form a cyclic group containing 3carbon atoms substituted with hydroxy, acyloxy, alkoxycarboxy,alkylthiocarboxy, hydroxymethyl, and aryloxycarboxy attached to a carbonatom that is three atoms from an oxygen attached to the phosphorus;

Z is selected from the group consisting of —CH₂OH, —CH₂OCOR³,—CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³, —S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar,—CH(Ar)OH, —CH(CH═CR²R²)OH, —CH(C≡CR²)OH, and —R²;

with the provisos that:

a) V, Z, W are not all —H; and

b) when Z is —R², then at least one of V and W is not —H or —R⁹;

R² is selected from the group consisting of R³ and —H;

R³ is selected from the group consisting of alkyl, aryl, alicyclic, andaralkyl; and

R⁹ is selected from the group consisting of alkyl, aralkyl, andalicyclic.

[9] Phosphoramidate derivatives have been explored as potentialphosphonate prodrugs (e.g. McGuigan et al., Antiviral Res. 1990, 14:345; 1991, 15: 255. Serafinowska et al., J. Med. Chem., 1995, 38,1372).Most phosphoramidates are unstable under aqueous acidic conditions andare hydrolyzed to the corresponding phosphonic acids. Cyclicphosphoramidates have also been studied as phosphonate prodrugs becauseof their potential for greater stability compared to non cyclicphosphoramidates (e.g. Starrett et al., J. Med. Chem., 1994, 37: 1857).

Other prodrugs are possible based on literature reports such assubstituted ethyls for example, bis(trichloroethyl)esters as disclosedby McGuigan, et al. Bioorg Med. Chem. Lett., 3:1207-1210 (1993), and thephenyl and benzyl combined nucleotide esters reported by Meier, C. etal. Bioorg. Med. Chem. Lett., 7:99-104 (1997).

X group nomenclature as used herein in formula 1 describes the groupattached to the phosphonate and ends with the group attached to the6-position of the purine ring. For example, when X is alkylamino, thefollowing structure is intended:

(OR¹)₂(O)P-alk-NR—(purine ring)

Y group nomenclature likewise ends with the group attached to the ring.

DETAILED DESCRIPTION OF THE INVENTION Novel Purine Compounds

Preferred compounds of the present invention are inhibitors of the AMPsite of FBPase of the following formula (1):

wherein

A is selected from the group consisting of —NR⁸ ₂, NHSO₂R³, —OR⁵, —SR⁵,halogen, lower alkyl, —CON(R⁴)₂, guanidine, amidine, —H, andperhaloalkyl;

E is selected from the group consisting of —H, halogen, lower alkylthio,lower perhaloalkyl, lower alkyl, lower alkenyl, lower alkynyl, loweralkoxy, —CN, and —NR⁷ ₂;

X is selected from the group consisting of alkylamino, alkyl, alkenyl,alkynyl, alkyl(carboxyl), alkyl(hydroxy), alkyl(phosphonate),alkyl(sulfonate), aryl, alkylaminoalkyl, alkoxyalkyl, alkylthioalkyl,alkylthio, alicyclic, 1,1-dihaloalkyl, carbonylalkyl,aminocarbonylamino, alkylaminocarbonyl, alkylcarbonylamino, aralkyl, andalkylaryl, all optionally substituted; or together with Y forms a cyclicgroup including cyclic alkyl, heterocyclic, and aryl;

Y is selected from the group consisting of —H, alkyl, alkenyl, alkynyl,aryl, alicyclic, aralkyl, aryloxyalkyl, alkoxyalkyl, —C(O)R³, —S(O)₂R³,—C(O)—OR³, —CONHR³, —NR² ₂, and —OR³, all except H are optionallysubstituted; or together with X forms a cyclic group including aryl,cyclic alkyl, and heterocyclic;

R¹ is independently selected from the group consisting of —H, alkyl,aryl, alicyclic where the cyclic moiety contains a carbonate orthiocarbonate, —C(R²)₂-aryl, alkylaryl, —C(R²)₂OC(O)NR² ₂, —NR²—C(O)—R³,—C(R²)₂—OC(O)R³, C(R²)₂—O—C(O)OR³, —C(R²)₂OC(O)SR³, alkyl-S—C(O)R³,alkyl-S—S-alkylhydroxy, and alkyl-S—S—S-alkylhydroxy, or together R¹ andR¹ are -alkyl-S—S-alkyl to form a cyclic group, or together R¹ and R¹are

wherein

V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or

together V and Z are connected to form a cyclic group containing 3-5atoms, optionally 1 heteroatom, substituted with hydroxy, acyloxy,alkoxycarboxy, or aryloxycarboxy attached to a carbon atom that is threeatoms from an oxygen attached to the phosphorus; or

together V and W are connected to form a cyclic group containing 3carbon atoms substituted with hydroxy, acyloxy, alkoxycarboxy,alkylthiocarboxy, hydroxymethyl, and aryloxycarboxy attached to a carbonatom that is three atoms from an oxygen attached to the phosphorus;

Z is selected from the group consisting of —CH₂OH, —C H₂OCOR³,—CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³, —S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar,—CH(Ar)OH, —CH(CH═CR²R²)OH, —CH(CH≡CR²)OH, and —R²;

with the provisos that:

a) V, Z, W are not all —H; and

b) when Z is —R², then at least one of V and W is not —H or —R⁹;

R² is selected from the group consisting of R³ and —H;

R³ is selected from the group consisting of alkyl, aryl, alicyclic, andaralkyl;

R⁴ is independently selected from the group consisting of —H, loweralkyl, lower alicyclic, lower aralkyl, and lower aryl;

R⁵ is selected from the group consisting of lower alkyl, lower aryl,lower aralkyl, and lower alicyclic;

R⁶ is independently selected from the group consisting of —H, and loweralkyl;

R⁷ is independently selected from the group consisting of —H, loweralkyl, lower alicyclic, lower aralkyl, lower aryl, and —C(O)R¹⁰;

R⁸ is independently selected from the group consisting of —H, loweralkyl, lower aralkyl, lower aryl, lower alicyclic, —C(O)R¹⁰, or togetherthey form a bidendate alkyl;

R⁹ is selected from the group consisting of alkyl, aralkyl, andalicyclic;

R¹⁰ is selected from the group consisting of —H, lower alkyl, —NH₂,lower aryl, and lower perhaloalkyl;

R¹¹ is selected from the group consisting of alkyl, aryl, —OH, —NH₂ and—OR³; and

pharmaceutically acceptable prodrugs and salts thereof.

Preferred Compounds of Formula 1

Suitable alkyl groups include groups having from 1 to about 20 carbonatoms. Suitable aryl groups include groups having from 1 to about 20carbon atoms. Suitable aralkyl groups include groups having from 2 toabout 21 carbon atoms. Suitable acyloxy groups include groups havingfrom 1 to about 20 carbon atoms. Suitable alkylene groups include groupshaving from 1 to about 20 carbon atoms. Suitable alicyclic groupsinclude groups having 3 to about 20 carbon atoms. Suitable heteroarylgroups include groups having from 1 to about 20 carbon atoms and from 1to 5 heteroatoms, preferably independently selected from nitrogen,oxygen, phosphorous, and sulfur. Suitable heteroalicyclic groups includegroups having from 2 to about twenty carbon atoms and from 1 to 5heteroatoms, preferably independently selected from nitrogen, oxygen,phosphorous, and sulfur.

Preferred A groups include —NR⁸ ₂, lower alkyl, lower perhaloalkyl,lower alkoxy, and halogen. Particularly preferred are —NR⁸ ₂, andhalogen. Especially preferred is —NR⁸ ₂. Most preferred is —NH₂.

Preferred E groups include —H, halogen, lower perhaloalkyl, —CN, loweralkyl, lower alkoxy, and lower alkylthio. Particularly preferred Egroups include —H, —SMe, —Et, and —Cl. Especially preferred is —H and—SCH₃.

Preferred X groups include alkylamino, alkyl, alkynyl, alkoxyalkyl,alkylthio, aryl, 1,1-dihaloalkyl, carbonylalkyl, heteroaryl,alkylcarbonylamino, and alkylaminocarbonyl. Particularly preferred isalkyl substituted with 1 to 3 substituents selected from halogen,phosphonate, —CO₂H, —SO₃H, and —OH. Particularly preferred arealkylaminocarbonyl, alkoxyalkyl, and heteroaryl. Preferred alkoxyalkylgroups include methoxymethyl. Preferred heteroaryl groups includefuranyl, optionally substituted.

Preferred Y groups include aralkyl, alicyclic, alkyl, and aryl, alloptionally substituted. Particularly preferred is lower alkyl.Particularly preferred Y groups include (2-naphthyl)methyl,cyclohexylethyl, phenylethyl, nonyl, cyclohexylpropyl, ethyl,cyclopropylmethyl, cyclobutylmethylphenyl, (2-methyl)propyl, neopentyl,cyclopropyl, cyclopentyl, (1-imidozolyl)propyl, 2-ethoxybenzyl,1-hydroxy-2,2-dimethylpropyl, 1-chloro-2,2-dimethylpropyl,2,2-dimethylbutyl , 2-(spiro-3,3-dimethylcyclohex-4-enyl)propyl, and1-methyineopentyl. Especially preferred is neopentyl and isobutyl.

Preferred R⁴ and R⁷ groups are —H, and lower alkyl. Particularlypreferred are —H, and methyl.

Preferred R¹ groups include —H, alkyl, aryl, alicyclic where the cyclicmoiety contains a carbonate or thiocarbonate, —C(R²)₂-aryl, alkylaryl,—C(R²)₂OC(O)NR² ₂, —NR²—C(O)—R³, —C(R²)₂—OC(O)R³, C(R²)₂—O—C(O)OR³,—C(R²)₂OC(O)SR³, alkyl-S—C(O)R³, alkyl-S—S-alkylhydroxy, andalkyl-S—S—S-alkylhydroxy, or together R¹ and R¹ are -alkyl-S—S-alkyl toform a cyclic group, or together R¹ and R¹ are

wherein

V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or

together V and Z are connected to form a cyclic group containing 3-5atoms, optionally 1 heteroatom, substituted with hydroxy, acyloxy,alkoxycarboxy, or aryloxycarboxy attached to a carbon atom that is threeatoms from an oxygen attached to the phosphorus; or

together V and W are connected to form a cyclic group containing 3carbon atoms substituted with hydroxy, acyloxy, alkoxycarboxy,alkylthiocarboxy, hydroxymethyl, and aryloxycarboxy attached to a carbonatom that is three atoms from an oxygen attached to the phosphorus;

Z is selected from the group consisting of —CH₂OH, —CH₂OCOR³,—CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³, —S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar,—CH(Ar)OH, —CH(CH═CR²R²)OH, —CH(C≡OCR²)OH, and —R²;

with the provisos that:

a) V, Z, W are not all —H; and

b) when Z is —R², then at least one of V and W is not —H or —R⁹;

R² is selected from the group consisting of R³ and —H;

R³ is selected from the group consisting of alkyl, aryl, alicyclic, andaralkyl; and

R⁹ is selected from the group consisting of alkyl, aralkyl, andalicyclic.

Preferred R¹ groups include —H, alkylaryl, aryl, —C(R²)₂-aryl, and—C(R²)₂ —OC(O)R³. Preferred such R¹ groups include optionallysubstituted phenyl, optionally substituted benzyl, —H, —C(R²)₂OC(O)OR³,and —C(R²)₂OC(O)R³. Preferably, said alkyl groups are greater than 4carbon atoms. Another preferred aspect is where at least one R¹ is arylor —C(R²)₂-aryl. Also particularly preferred are compounds where R¹ isalicyclic where the cyclic moiety contains carbonate or thiocarbonate.Another preferred aspect is when at least one R¹ is —C(R²)₂—OC(O)R³,—C(R²)₂—OC(O)OR³ or —C(R²)₂—OC(O)SR³. Also particularly preferred iswhen R¹ and R¹ together are optionally substituted, including fused,lactone attached at the omega position or are optionally substituted2-oxo-1,3-dioxolenes attached through a methylene to the phosphorusoxygen. Also preferred is when at least one R¹ is-alkyl-S—S-alkylhydroxyl, -alkyl-S—C(O)R³, and-alkyl-S—S—S-alkylhydroxy, or together R¹ and R¹ are -alkyl-S—S-alkyl-to form a cyclic group. Also preferred is where R¹ and R¹ together are

to form a cyclic group,

wherein

V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or

together V and Z are connected to form a cyclic group containing 3-5atoms, optionally 1 heteroatom, substituted with hydroxy, acyloxy,alkoxycarboxy, or aryloxycarboxy attached to a carbon atom that is threeatoms from an oxygen attached to the phosphorus; or

together V and W are connected to form a cyclic group containing 3carbon atoms substituted with hydroxy, acyloxy, alkoxycarboxy,alkylthiocarboxy, hydroxymethyl, and aryloxycarboxy attached to a carbonatom that is three atoms from an oxygen attached to the phosphorus;

Z is selected from the group consisting of —CH₂OH, —CH₂OCOR³,—CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³, —S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar,—CH(Ar)OH, —CH(CH═CR²R²)OH, —CH(C≡CR²)OH, and —R²;

with the provisos that:

a) V, Z, W are not all —H; and

b) when Z is —R², then at least one of V and W is not —H or —R⁹;

R² is selected from the group consisting of R³ and —H;

R³ is selected from the group consisting of alkyl, aryl, alicyclic, andaralkyl; and

R⁹ is selected from the group consisting of alkyl, aralkyl, andalicyclic.

Particularly preferred are such groups wherein V and W both form a6-membered carbocyclic ring substituted with 0-4 groups, selected fromthe group consisting of hydroxy, acyloxy, alkoxycarbonyl, and alkoxy;and Z is R². Also particularly preferred are such groups wherein V and Ware hydrogen; and Z is selected from the group consisting ofhydroxyalkyl, acyloxyalkyl, alkyloxyalkyl, and alkoxycarboxyalkyl. Alsoparticularly preferred are such groups wherein V and W are independentlyselected from the group consisting of hydrogen, optionally substitutedaryl, and optionally substituted heteroaryl, with the proviso that atleast one of V and W is optionally substituted aryl or optionallysubstituted heteroaryl.

In one preferred aspect, R¹ is not lower alkyl of 1-4 carbon atoms.

In another preferred aspect, A is —NR⁸ ₂ or halogen, E is —H, halogen,—CN, lower alkyl, lower perhaloalkyl, lower alkoxy, or lower alkylthio,X is alkylamino, alkyl, alkoxyalkyl, alkynyl, 1,1-dihaloalkyl,carbonylakyl, alkyl(OH), alkyl(sulfonate), alkylcarbonylamino,alkylaminocarbonyl, alkylthio, aryl, or heteroaryl, and R⁴ and R⁷ is —Hor lower alkyl. Particularly preferred are such compounds where Y isaralkyl, aryl, alicyclic, or alkyl. Especially preferred are suchcompounds where R¹ and R¹ together are

wherein

V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or

together V and Z are connected to form a cyclic group containing 3-5atoms, optionally 1 heteroatom, substituted with hydroxy, acyloxy,alkoxycarboxy, or aryloxycarboxy attached to a carbon atom that is threeatoms from an oxygen attached to the phosphorus; or

together V and W are connected to form a cyclic group containing 3carbon atoms substituted with hydroxy, acyloxy, alkoxycarboxy,alkylthiocarboxy, hydroxymethyl, and aryloxycarboxy attached to a carbonatom that is three atoms from an oxygen attached to the phosphorus;

Z is selected from the group consisting of —CH₂OH, —CH₂OCOR³,—CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³, —S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar,—CH(Ar)OH, —CH(CH═CR²R²)OH, —CH(C≡CR²)OH, and —R²;

with the provisos that:

a) V, Z, W are not all —H; and

b) when Z is —R², then at least one of V and W is not —H or —R⁹;

R² is selected from the group consisting of R³ and —H;

R³ is selected from the group consisting of alkyl, aryl, alicyclic, andaralkyl; and

R⁹ is selected from the group consisting of alkyl, aralkyl, andalicyclic.

In another preferred aspect, A is —NR⁸ ₂, E is —H, Cl—, or methylthio,and X is optionally substituted furanyl, or alkoxyalkyl. Particularlypreferred are such compounds where A is —NH₂, X is 2,5-furanyl, ormethoxymethyl, and Y is lower alkyl. Most preferred are such compoundswhere E is H, X is 2,5-furanyl, and Y is neopentyl; those where E is—SCH₃, X is 2,5-furanyl, and Y is isobutyl; and those where E is —H, Xis 2,5-furanyl, and Y is 1-(3-chloro-2,2-dimethyl)-propyl. Especiallypreferred are such compounds where R¹ is —CH₂O—C(O)—C(CH₃)₃.

In the following examples of preferred compounds, the following prodrugsare preferred:

Acyloxyalkyl esters;

Alkoxycarbonyloxyalkyl esters;

Aryl esters;

Benzyl and substituted benzyl esters;

Disulfide containing esters;

Substituted (1,3-dioxolen-2-one)methyl esters;

Substituted 3-phthalidyl esters;

Cyclic-[2′-hydroxymethyl]-1,3-propanyl diesters and hydroxy protectedforms;

Lactone type esters; and all mixed esters resulted from possiblecombinations of above esters.

Bis-pivaloyloxymethyl esters;

Bis-isobutyryloxymethyl esters;

Cyclic-[2′-hydroxymethyl]-1,3-propanyl diester;

Cyclic-[2′-acetoxymethyl]-1,3-propanyl diester;

Cyclic-[2′-methyloxycarbonyloxymethyl]-1,3-propanyl diester;

Bis-benzoylthiomethyl esters;

Bis-benzoylthioethyl esters;

Bis-benzoyloxymethyl esters;

Bis-p-fluorobenzoyloxymethyl esters;

Bis-6-chloronicotinoyloxymethyl esters;

Bis-5-bromonicotinoyloxymethyl esters;

Bis-thiophenecarbonyloxymethyl esters;

Bis-2-furoyloxymethyl esters;

Bis-3-furoyloxymethyl esters;

Diphenyl esters;

Bis-(4-methoxyphenyl) esters;

Bis-(2-methoxyphenyl) esters;

Bis-(2-ethoxyphenyl) esters;

Mono-(2-ethoxyphenyl) esters;

Bis-(4-acetamidophenyl) esters;

Bis-(4-acetoxyphenyl) esters;

Bis-(4-hydroxyphenyl) esters;

Bis-(2-acetoxyphenyl) esters;

Bis-(3-acetoxyphenyl) esters;

Bis-(4-morpholinophenyl) esters;

Bis-[4-(1-triazolophenyl) esters;

Bis-(3-N,N-dimethylaminophenyl) esters;

Bis-(2-tetrahydronapthyl) esters;

Bis-(3-chloro-4-methoxy)benzyl esters;

Bis-(3-bromo-4-methoxy)benzyl esters;

Bis-(3-cyano-4-methoxy)benzyl esters;

Bis-(3-chloro-4-acetoxy)benzyl esters;

Bis-(3-bromo-4-acetoxy)benzyl esters;

Bis-(3-cyano-4-acetoxy)benzyl esters;

Bis-(4-chloro)benzyl esters;

Bis-(4-acetoxy)benzyl esters;

Bis-(3,5-dimethoxy-4-acetoxy)benzyl esters;

Bis-(3-methyl-4-acetoxy)benzyl esters;

Bis-(benzyl)esters;

Bis-(3-methoxy-4-acetoxy)benzyl esters;

Bis-(3-chloro-4-acetoxy)benzyl esters;

cyclic-(2,2-dimethylpropyl)phosphonoamidate;

cyclic-(2-hydroxymethylpropyl) ester;

Bis-(6′-hydroxy-3′,4′-disulfide)hexyl esters;

Bis-(6′-acetoxy-3′,4′-disulfide)hexyl esters;

(3′,4′-Dithia)cyclononane esters;

Bis-(5-methyl-1,3-dioxolen-2-one-4-yl)methyl esters;

Bis-(5-ethyl-1,3-dioxolen-2-one-4-yl)methyl esters;

Bis-(5-tert-butyl-1,3-dioxolen-2-one-4-yl)methyl esters;

Bis-3-(5,6,7-trimethoxy)phthalidyl esters;

Bis-(cyclohexyloxycarbonyloxymethyl) esters;

Bis-(isopropyloxycarbonyloxymethyl) esters;

Bis-(ethyloxycarbonyloxymethyl) esters;

Bis-(methyloxycarbonyloxymethyl) esters;

Bis-(isopropylthiocarbonyloxymethyl) esters;

Bis-(phenyloxycarbonyloxymethyl) esters;

Bis-(benzyloxycarbonyloxymethyl) esters;

Bis-(phenylthiocarbonyloxymethyl) esters;

Bis-(p-methoxyphenyloxycarbonyloxymethyl) esters;

Bis-(m-methoxyphenyloxycarbonyloxymethyl) esters;

Bis-(o-methoxyphenyloxycarbonyloxymethyl) esters;

Bis-(o-methylphenyloxycarbonyloxymethyl) esters;

Bis-(p-chlorophenyloxycarbonyloxymethyl) esters;

Bis-(1,4-biphenyloxycarbonyloxymethyl) esters;

Bis-[(2-phthalimidoethyl)oxycarbonyloxymethyl]esters;

Bis-(N-Phenyl, N-methylcarbamoyloxymethyl) esters;

Bis-(2-trichloroethyl) esters;

Bis-(2-bromoethyl) esters;

Bis-(2-iodoethyl) esters;

Bis-(2-azidoethyl) esters;

Bis-(2-acetoxyethyl) esters;

Bis-(2-aminoethyl) esters;

Bis-(2-N,N-diaminoethyl) esters;

Bis-(2-aminoethyl) esters;

Bis-(methoxycarbonylmethyl) esters;

Bis-(2-aminoethyl) esters;

Bis-[N,N-di(2-hydroxyethyl)]amidomethylesters;

Bis-(2-aminoethyl) esters;

Bis-(2-methyl-5-thiozolomethyl) esters;

Bis-(bis-2-hydroxyethylamidomethyl) esters.

Most preferred are the following:

Bis-pivaloyloxymethyl esters;

Bis-isobutyryloxymethyl esters;

cyclic-(2-hydroxymethylpropyl) ester;

cyclic-(2-acetoxymethylpropyl) ester;

cyclic-(2-methyloxycarbonyloxymethylpropyl) ester;

cyclic-(2-cyclohexylcarbonyloxymethylpropyl)ester;

cyclic-(2-aminomethylpropyl)ester;

cyclic-(2-azidomethylpropyl)ester;

Bis-benzoylthiomethyl esters;

Bis-benzoylthioethylesters;

Bis-benzoyloxymethyl esters;

Bis-p-fluorobenzoyloxymethyl esters;

Bis-6-chloronicotinoyloxymethyl esters;

Bis-5-bromonicotinoyloxymethyl esters;

Bis-thiophenecarbonyloxymethyl esters;

Bis-2-furoyloxymethyl esters;

Bis-3-furoyloxymethyl esters;

Diphenyl esters;

Bis-(2-methyl)phenyl esters;

Bis-(2-methoxy)phenyl esters;

Bis-(2-ethoxy)phenyl esters;

Bis-(4-methoxy)phenyl esters;

Bis-(3-bromo-4-methoxy)benzyl esters;

Bis-(4-acetoxy)benzyl esters;

Bis-(3,5-dimethoxy-4-acetoxy)benzyl esters;

Bis-(3-methyl-4-acetoxy)benzyl esters;

Bis-(3-methoxy-4-acetoxy)benzyl esters;

Bis-(3-chloro-4-acetoxy)benzyl esters;

Bis-(cyclohexyloxycarbonyloxymethyl) esters;

Bis-(isopropyloxycarbonyloxymethyl) esters;

Bis-(ethyloxycarbonyloxymethyl) esters;

Bis-(methyloxycarbonyloxymethyl) esters;

Bis-(isopropylthiocarbonyloxymethyl) esters;

Bis-(phenyloxycarbonyloxymethyl) esters;

Bis-(benzyloxycarbonyloxymethyl) esters;

Bis-(phenylthiocarbonyloxymethyl) esters;

Bis-(p-methoxyphenyloxycarbonyloxymethyl) esters;

Bis-(m-methoxyphenyloxycarbonyloxymethyl) esters;

Bis-(o-methoxyphenyloxycarbonyloxymethyl) esters;

Bis-(o-methylphenyloxycarbonyloxymethyl) esters;

Bis-(p-chlorophenyloxycarbonyloxymethyl) esters;

Bis-(1,4-biphenyloxycarbonyloxymethyl) esters;

Bis-[(2-phthalimidoethyl)oxycarbonyloxymethyl]esters;

Bis-(6′-hydroxy-3′,4′-disulfide)hexyl esters; and

(3′,4′-Disulfide)cyclononane esters.

Bis-(2-bromoethyl) esters;

Bis-(2-aminoethyl) esters;

Bis-(2-N,N-diaminoethyl) esters;

Examples of preferred compounds include, but are not limited to thosedescribed in Table 1 including salts and prodrugs thereof:

Table Compound No. Synthetic Example No. A E

Y X 1 2.7 NH2 H neopentyl 2,5-furanyl 2 H H neopentyl 2,5-furanyl 3 Me Hneopentyl 2,5-furanyl 4 Et H neopentyl 2,5-furanyl 5 Pr H neopentyl2,5-furanyl 6 I H neopentyl 2,5-furanyl 7 Br H neopentyl 2,5-furanyl 8Cl H neopentyl 2,5-furanyl 9 F H neopentyl 2,5-furanyl 10 OMe Hneopentyl 2,5-furanyl 11 OEt H neopentyl 2,5-furanyl 12 OPr H neopentyl2,5-furanyl 13 SMe H neopentyl 2,5-furanyl 14 SEt H neopentyl2,5-furanyl 15 SPr H neopentyl 2,5-furanyl 16 SBn H neopentyl2,5-furanyl 17 OBn H neopentyl 2,5-furanyl 18 NHMe H neopentyl2,5-furanyl 19 NHEt H neopentyl 2,5-furanyl 20 NH-cPr H neopentyl2,5-furanyl 21 NHOH H neopentyl 2,5-furanyl 22 NHNH2 H neopentyl2,5-furanyl 23 NHCHO H neopentyl 2,5-furanyl 24 NHAc H neopentyl2,5-furanyl 25 NHCOCF3 H neopentyl 2,5-furanyl 26 NHSO2Me H neopentyl2,5-furanyl 27 CONH2 H neopentyl 2,5-furanyl 28 ONH2 H neopentyl2,5-furanyl 29 22.2 NH2 SMe neopentyl 2,5-furanyl 30 H SMe neopentyl2,5-furanyl 31 Me SMe neopentyl 2,5-furanyl 32 Et SMe neopentyl2,5-furanyl 33 Pr SMe neopentyl 2,5-furanyl 34 I SMe neopentyl2,5-furanyl 35 Br SMe neopentyl 2,5-furanyl 36 Cl SMe neopentyl2,5-furanyl 37 F SMe neopentyl 2,5-furanyl 38 OMe SMe neopentyl2,5-furanyl 39 OEt SMe neopentyl 2,5-furanyl 40 OPr SMe neopentyl2,5-furanyl 41 SMe SMe neopentyl 2,5-furanyl 42 SEt SMe neopentyl2,5-furanyl 43 SPr SMe neopentyl 2,5-furanyl 44 SBn SMe neopentyl2,5-furanyl 45 OBn SMe neopentyl 2,5-furanyl 46 NHMe SMe neopentyl2,5-furanyl 47 NHEt SMe neopentyl 2,5-furanyl 48 NH-cPr SMe neopentyl2,5-furanyl 49 NHOH SMe neopentyl 2,5-furanyl 50 NHNH2 SMe neopentyl2,5-furanyl 51 NHCHO SMe neopentyl 2,5-furanyl 52 NHAc SMe neopentyl2,5-furanyl 53 NHCOCF3 SMe neopentyl 2,5-furanyl 54 NHSO2Me SMeneopentyl 2,5-furanyl 55 CONH2 SMe neopentyl 2,5-furanyl 56 ONH2 SMeneopentyl 2,5-furanyl 57 NH2 H isobutyl 2,5-furanyl 58 2.6 NH2 Hisopropyl 2,5-furanyl 59 2.5 NH2 H ethyl 2,5-furanyl 60 NH2 H methyl2,5-furanyl 61 2.9 NH2 H cyclopropyl 2,5-furanyl 62 NH2 H cyclobutyl2,5-furanyl 63 2.10 NH2 H cyclopentyl 2,5-furanyl 64 NH2 H cyclohexyl2,5-furanyl 65 NH2 H cycloheptanyl 2,5-furanyl 66 NH2 Hcyclopropylmethyl 2,5-furanyl 67 NH2 H cyclobutylmethyl 2,5-furanyl 68NH2 H cyclopentylmethyl 2,5-furanyl 69 NH2 H 2-cyclopropylethyl2,5-furanyl 70 NH2 H 2-cyclobutylethyl 2,5-furanyl 71 NH2 H2-cyclopentylethyl 2,5-furanyl 72 2.2 NH2 H 2-cyclohexylethyl2,5-furanyl 73 2.1 NH2 H 2-phenylethyl 2,5-furanyl 74 NH2 H benzyl2,5-furanyl 75 NH2 H phenyl 2,5-furanyl 76 NH2 H D-ribosyl 2,5-furanyl77 NH2 H H 2,5-furanyl 78 NH2 H 1-naphthylmethyl 2,5-furanyl 79 2.3 NH2H 2-naphthylmethyl 2,5-furanyl 80 NH2 H 3-cyclopropylpropyl 2,5-furanyl81 NH2 H 3-cyclobutylpropyl 2,5-furanyl 82 NH2 H 3-cyclopentylpropyl2,5-furanyl 83 NH2 H 3-cyclohexylpropyl 2,5-furanyl 84 2.4 NH2 H2,2-diphenylethyl 2,5-furanyl 85 2.8 NH2 H adamentylmethyl 2,5-furanyl86 2.11 NH2 H 2-ethoxybenzyl 2,5-furanyl 87 2.13 NH2 H 2,2-dimethyl-3-2,5-furanyl hydroxy-1-propyl 88 2.12 NH2 H 2,2-dimethyl-3- 2,5-furanyldimethylamino-1- propyl 89 2.14 NH2 H 2,2-dimethyl-3- 2,5-furanylchloro-1-propyl 90 2.15 NH2 H 3,3-dimethyl-1-butyl 2,5-furanyl 91 2.17NH2 H 1,2,2-trimethyl-1- 2,5-furanyl propyl 92 2.16 NH2 H1,5,5-trimethyl-3-ene- 2,5-furanyl 1-cyclohexylmethyl 93 NH2 H4-pyrimidylmethyl 2,5-furanyl 94 NH2 H 2-(4-pyrimidyl)ethyl 2,5-furanyl95 NH2 H 5-pyrimidylmethyl 2,5-furanyl 96 NH2 H 2-(5-pyrimidyl)ethyl2,5-furanyl 97 NH2 H 2-pyrimidylmethyl 2,5-furanyl 98 NH2 H2-(2-pyrimidyl)ethyl 2,5-furanyl 99 NH2 H 2-pyridylmethyl 2,5-furanyl100 NH2 H 2-(2-pyridyl)ethyl 2,5-furanyl 101 NH2 H 3-pyridylmethyl2,5-furanyl 102 NH2 H 2-(3-pyridyl)ethyl 2,5-furanyl 103 NH2 H4-pyridylmethyl 2,5-furanyl 104 NH2 H 2-(4-pyridyl)ethyl 2,5-furanyl 105NH2 H 2-carbamoylethyl 2,5-furanyl 106 NH2 H 1-(2- 2,5-furanylcarbamoyl)propyl 107 NH2 H neopentyl CONHCH2 108 NH2 H neopentylCONHCH2CH2 109 NH2 H neopentyl CH2CH2CH2 110 NH2 H neopentyl CH2CH2CF2111 5.5 NH2 H neopentyl CH2OCH2 112 NH2 H neopentyl CH2OCF2 113 NH2 Hneopentyl CF2CF2CF2 114 NH2 H neopentyl acetylene 115 NH2 H neopentylSCH2 116 NH2 H neopentyl SCH2CH2 117 NH2 H neopentyl CH2SCH2 118 NH2 Hneopentyl NHCH2CH2 119 NH2 H neopentyl N(Ac)CH2CH2 120 NH2 H neopentylN(Bz)CH2CH2 121 NH2 H neopentyl N(Me)CH2CH2 122 NH2 H neopentylN(Bn)CH2CH2 123 NH2 H neopentyl NHCOCH2 124 NH2 H neopentyl NHCOCH2CH2125 NH2 H neopentyl NHCOCF2 126 NH2 H neopentyl NHSO2CH2 127 NH2 Hneopentyl N(Me)COCH2 128 NH2 H neopentyl N(Bn)COCH2 129 NH2 H neopentylNHOCH2 130 NH2 H neopentyl CH2CH2CH(OH) 131 NH2 H neopentylCH2CH2CH(CO2H) 132 NH2 H neopentyl CH2CH2CH(SO3H) 133 NH2 H neopentylCH2CH2CH(PO3H2) 134 20.1 NH2 H neopentyl CH2-(1,2-imidazyl) 135 NH2 Hneopentyl CH2-(1,2-pyrrolyl) 136 NH2 H neopentyl CSNHCH2 137 NH2 Hneopentyl 2,5- tetrahydrofuranyl 138 NH2 H neopentyl 2,5-pyrrolidinyl139 NH2 H neopentyl 3,4-dihydroxy-2,5- tetrahydrofuranyl 140 NH2 Hneopentyl 2,4-furanyl 141 NH2 H neopentyl 4,2-furanyl 142 NH2 Hneopentyl 2,5-thienyl 143 NH2 H neopentyl 2,4-thienyl 144 NH2 Hneopentyl 4,2-thienyl 145 NH2 H neopentyl 2,5-pyrrolyl 146 NH2 Hneopentyl 2,5-imidazyl 147 NH2 H neopentyl 5,2-imidazyl 148 NH2 Hneopentyl 2,5-oxazyl 149 NH2 H neopentyl 5,2-oxazyl 150 12.1 NH2 Hneopentyl 3,4-dichloro-2,5- furanyl 151 NH2 H neopentyl 3-chloro-2,5-furanyl 152 NH2 H neopentyl 4-chloro-2,5- furanyl 153 NH2 H neopentyl3,4-fluoro-2,5- furanyl 154 NH2 H neopentyl 3-fluoro-2,5-furanyl 155 NH2H neopentyl 4-fluoro-2,5-furanyl 156 NH2 H neopentyl CONHCH(CO2H) 157NH2 Me neopentyl 2,5-furanyl 158 NH2 Et neopentyl 2,5-furanyl 159 NH2 Prneopentyl 2,5-furanyl 160 NH2 vinyl neopentyl 2,5-furanyl 161 NH2acetylen neopentyl 2,5-furanyl yl 162 NH2 allyl neopentyl 2,5-furanyl163 NH2 2- neopentyl 2,5-furanyl furanyl 164 NH2 3- neopentyl2,5-furanyl furanyl 165 NH2 2- neopentyl 2,5-furanyl thienyl 166 NH2 3-neopentyl 2,5-furanyl thienyl 167 NH2 Ph neopentyl 2,5-furanyl 168 22.1NH2 NH2 neopentyl 2,5-furanyl 169 NH2 NHMe neopentyl 2,5-furanyl 170 NH2N(Me)2 neopentyl 2,5-furanyl 171 NH2 NHBn neopentyl 2,5-furanyl 172 NH2I neopentyl 2,5-furanyl 173 NH2 Br neopentyl 2,5-furanyl 174 NH2 Clneopentyl 2,5-furanyl 175 NH2 F neopentyl 2,5-furanyl 176 NH2 OMeneopentyl 2,5-furanyl 177 NH2 OEt neopentyl 2,5-furanyl 178 NH2 OPrneopentyl 2,5-furanyl 179 NH2 SO2Me neopentyl 2,5-furanyl 180 NH2 SEtneopentyl 2,5-furanyl 181 NH2 SPr neopentyl 2,5-furanyl 182 NH2 SBuneopentyl 2,5-furanyl 183 NH2 CN neopentyl 2,5-furanyl 184 NH2 CONH2neopentyl 2,5-furanyl 185 NH2 2- neopentyl 2,5-furanyl pyridyl 186 NH23- neopentyl 2,5-furanyl pyridyl 187 NH2 4- neopentyl 2,5-furanylpyridyl 188 5.4 NH2 H 1-(3- CH2OCH2 cyclohexyl)propyl 189 5.3 NH2 H1-nonyl CH2OCH2 190 5.2 NH2 H 2-cyclohexylethyl CH2OCH2 191 5.1 NH2 H2-phenethyl CH2OCH2 192 10.2 NHMe H 2-phenethyl 2,5-furanyl 193 10.1N(Me)2 H 2-phenethyl 2,5-furanyl 194 9.1 Cl H 2-phenethyl 2,5-furanyl195 11.1 NH2 SMe isobutyl 2,5-furanyl 196 11.2 NH2 SO2Me isobutyl2,5-furanyl 197 4.1 NH2 H D-ribosyl NHCH2CH2 198 4.2 NH2 H5′-deoxy-D-ribosyl NHCH2CH2 199 NH2 H H NHCH2CH2 200 3.1 NH2 H benzylNHCH2CH2 201 3.2 NH2 H 2-phenethyl NHCH2CH2 202 3.3 NH2 H2-naphthylmethyl NHCH2CH2 203 6.2 NH2 H 2-phenethyl CH2CH2CH2 204 3.4NH2 H 2-cyclohexylethyl NHCH2CH2 205 6.1 NH2 H 2-cyclohexylethylCH2CH2CH2 206 8.1 NH2 H 2-cyclohexylethyl SCH2 207 7.1 NH2 H 2-phenethyl2,5-thienyl 208 NH2 Me isobutyl 2,5-furanyl 209 NH2 Et isobutyl2,5-furanyl 210 NH2 SEt isobutyl 2,5-furanyl 211 NH2 SPr isobutyl2,5-furanyl 212 NH2 2- isobutyl 2,5-furanyl furanyl 213 NH2 2- isobutyl2,5-furanyl thienyl 214 NH2 Pr isobutyl 2,5-furanyl 215 NH2 F isobutyl2,5-furanyl 216 NH2 Cl isobutyl 2,5-furanyl 217 NH2 Br isobutyl2,5-furanyl 218 NH2 H isobutyl 2,5-furanyl 219 NH2 Et isobutyl CONHCH2220 NH2 SEt isobutyl CONHCH2 221 NH2 SPr isobutyl CONHCH2 222 NH2 2-isobutyl CONHCH2 furanyl 223 NH2 2- isobutyl CONHCH2 thienyl 224 NH2 Prisobutyl CONHCH2 225 NH2 F isobutyl CONHCH2 226 NH2 Cl isobutyl CONHCH2227 NH2 Br isobutyl CONHCH2 228 NH2 Me isobutyl CONHCH2 229 NH2 Hisobutyl CONHCH2 230 NH2 Et neopentyl acetylene 231 NH2 SEt neopentylacetylene 232 NH2 SPr neopentyl acetylene 233 NH2 2- neopentyl acetylenefuranyl 234 NH2 2- neopentyl acetylene thienyl 235 NH2 Pr neopentylacetylene 236 NH2 F neopentyl acetylene 237 NH2 Cl neopentyl acetylene238 NH2 Br neopentyl acetylene 239 NH2 Me neopentyl acetylene 240 NH2 Etneopentyl NHCOCH2 241 NH2 SEt neopentyl NHCOCH2 242 NH2 SPr neopentylNHCOCH2 243 NH2 2- neopentyl NHCOCH2 furanyl 244 NH2 2- neopentylNHCOCH2 thienyl 245 NH2 Pr neopentyl NHCOCH2 246 NH2 F neopentyl NHCOCH2247 NH2 Cl neopentyl NHCOCH2 248 NH2 Br neopentyl NHCOCH2 249 NH2 Meneopentyl NHCOCH2 250 NH2 Et neopentyl CH2OCH2 251 NH2 SEt neopentylCH2OCH2 252 NH2 SPr neopentyl CH2OCH2 253 NH2 2- neopentyl CH2OCH2furanyl 254 NH2 2- neopentyl CH2OCH2 thienyl 255 NH2 Pr neopentylCH2OCH2 256 NH2 F neopentyl CH2OCH2 257 NH2 Cl neopentyl CH2OCH2 258 NH2Br neopentyl CH2OCH2 259 NH2 Me neopentyl CH2OCH2 260 NHBn H neopentyl2,5-furanyl 261 NHPh H neopentyl 2,5-furanyl 262 NHBn SMe neopentyl2,5-furanyl 263 NHPh SMe neopentyl 2,5-furanyl 264 NHPh-4-F H neopentyl2,5-furanyl 265 NHPh-4-F SMe neopentyl 2,5-furanyl 266 NHNH2 F neopentyl2,5-furanyl 267 NH2 Me cyclopropylmethyl 2,5-furanyl 268 NH2 SMecyclopropylmethyl 2,5-furanyl 269 NH2 F cyclopropylmethyl 2,5-furanyl270 NH2 Cl cyclopropylmethyl 2,5-furanyl 271 NH2 Br cyclopropylmethyl2,5-furanyl 272 NH2 Et cyclopropylmethyl 2,5-furanyl 273 NH2 CNcyclopropylmethyl 2,5-furanyl 274 NH2 Me cyclopropylmethyl CONHCH2 275NH2 SMe cyclopropylmethyl CONHCH2 276 NH2 F cyclopropylmethyl CONHCH2277 NH2 Cl cyclopropylmethyl CONHCH2 278 NH2 Br cyclopropylmethylCONHCH2 279 NH2 Et cyclopropylmethyl CONHCH2 280 NH2 CNcyclopropylmethyl CONHCH2 281 NH2 Me cyclopropylmethyl NHCOCH2 282 NH2SMe cyclopropylmethyl NHCOCH2 283 NH2 F cyclopropylmethyl NHCOCH2 284NH2 Cl cyclopropylmethyl NHCOCH2 285 NH2 Br cyclopropylmethyl NHCOCH2286 NH2 Et cyclopropylmethyl NHCOCH2 287 NH2 CN cyclopropylmethylNHCOCH2 288 2.18 NH2 H 3-(1-imidazolylpropyl) 2,5-furanyl 289 19.1 NH2 Hneopentyl 1,2-C₆H₄—O— 290 21.1 NH2 H 2-phenethyl CONHCH2

More preferred are the following compounds from Table 1 including saltsand prodrugs thereof:

1, 21, 22, 23, 29, 50, 57, 61, 62, 63, 66, 67, 72, 73, 89, 90, 107, 110,111, 112, 113, 114, 115, 119, 123, 125, 126, 129, 130, 131, 132, 133,134, 136, 137, 145, 148, 149, 151, 152, 153, 154, 155, 156, 158, 159,163, 165, 173, 174, 175, 180, 181, 182, 183, 209, 210, 212, 215, 216,217, 219, 220, 221, 230, 231, 234, 236, 237, 238, 240, 241, 246, 247,248, 250, 251, 256, 257, 258, 266, 268, 269, and 272.

Most preferred are the following compounds and their salts and prodrugs:

N⁹-neopentyl-2-methylthio-8-phosphonomethylaminocarbonyladenine;

N⁹-neopentyl-2-methylthio-8-(2-(5-phosphono)furanyl)adenine;

N⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine;

N⁹-isobutyl-2-methylthio-8-(2-(5-phosphono)furanyl)adenine;

N⁹-isobutyl-8-(2-(5-phosphono)furanyl)adenine;

N⁹-cyclopropyl-8-(2-(5-phosphono)furanyl)adenine;

N⁹-(2-cyclohexyl)ethyl-8-(2-(5-phosphono)furanyl)adenine;

N⁹-(1-(2,2-dimethyl-3-chloro)propyl)-8-(2-(5-phosphono)furanyl)adenine;

N⁹-(1-(3,3-dimethyl)butyl)-8-(2-(5-phosphono)furanyl)adenine;

N⁹-(1,5,5-trimethyl-3-cyclohexen-1-yl)methyl-8-(2-(5-phosphono)furanyl)adenine;

N⁹-neopentyl-8-(2-phosphonoacetylene-1-yl)adenine;

N⁹-neopentyl-8-(1-(3-phosphono-3-sulfuryl)propyl)adenine;

N⁹-neopentyl-8-(1-(3-phosphono-3-carboxyl)propyl)adenine;

N⁹-neopentyl-8-(1-(3,3-diphosphono)propyl)adenine;

N⁹-neopentyl-2-chloro-8-(2-(5-phosphono)furanyl)adenine;

2-Ethyl-N⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine;

2-Methylthio-N⁹-isobutyl-8-(2-(5-phosphono)furanyl)adenine; and

2-Methylthio-N⁹-isobutyl-8-(phosphonomethoxymethyl)adenine.

Synthesis of Compounds of Formula 1

Synthesis of compounds encompassed by the present invention typicallyincludes some or all of the following general steps: (1) preparation ofphosphonate prodrug; (2) deprotection of phosphonate ester; (3)modification of C-8-substituted purine intermediates; (4) modificationof purine at positions other than C-8; (5) construction of the purinering system; and (6) preparation of 4,5-diaminopyrimidine and othercoupling partners.

(1) Preparation of Phosphonate Prodrugs

Prodrug esters can be introduced at different stages of the synthesis.

Because of their lability, prodrugs are often prepared from compounds offormula 1 where R¹ is H. Advantageously, these prodrug esters can beintroduced at an early stage, provided that it can withstand thereaction conditions of the subsequent steps.

Compounds of formula 5 where R¹ is H, can be alkylated withelectrophiles (such as alkyl halides, alkyl sulfonates etc) undernucleophilic substitution reaction conditions to give phosphonateesters. For example prodrugs of formula 1, where R¹ is acyloxymethylgroup can be synthesized through direct alkylation of the freephosphonic acid of formula 5, with the desired acyloxymethyl halide(e.g. Cl, Br, I; Elhaddadi, et al Phosphorus Sulfur, 1990, 54(1-4): 143;Hoffmann, Synthesis, 1988, 62) in presence of base e.g. N,N′-dicyclohexyl-4-morpholinecarboxamidine, Hunigs base etc. in polaraprotic solvents such as DMF (Starrett, et al, J. Med. Chem., 1994,1857). These carboxylates include but not limited to acetate, propylate,isobutyrate, pivalate, benzoate, and other carboxylates. Alternately,these acyloxymethylphosphonate esters can also be synthesized bytreatment of the nitrophosphonic acid (A is NO₂ in formula 5; Dickson,et al, J. Med. Chem., 1996, 39: 661; lyer, et al, Tetrahedron Lett.,1989, 30: 7141; Srivastva, et al, Bioorg. Chem., 1984, 12: 118). Thiscan be extended to many other types of prodrugs, such as compounds offormula 1 where R¹ is 3-phthalidyl,2-oxo-4,5-didehydro-1,3-dioxolanemethyl, and 2-oxotetrahydrofuran-5-ylgroups, etc. (Biller and Magnin (U.S. Pat. No. 5,157,027); Serafinowskaet al. (J. Med. Chem. 38: 1372 (1995)); Starrett et al. (J. Med. Chem.37: 1857 (1994)); Martin et al. J. Pharm. Sci. 76: 180 (1987); Alexanderet al., Collect. Czech. Chem. Commun, 59: 1853 (1994)); and EPO0632048A1). N,N-Dimethylformamide dialkyl acetals can also be used toalkylate phosphonic acids (Alexander, P., et al Collect. Czech. Chem.Commun., 1994, 59,1853).

Alternatively, these phosphonate prodrugs or phosphoramidates can alsobe synthesized, by reaction of the corresponding dichlorophosphonatesand an alcohol or an amine (Alexander, et al, Collect. Czech. Chem.Commun., 1994, 59: 1853). For example, the reaction ofdichlorophosphonate with phenols and benzyl alcohols in the presence ofbase (such as pyridine, triethylamine, etc) yields compounds of formula1 where R¹ is aryl (Khamnei, S., et al J. Med. Chem., 1996, 39: 4109;Serafinowska, H. T., et al J. Med. Chem., 1995, 38:1372; De Lombaert,S., et al J. Med. Chem., 1994, 37: 498) or benzyl (Mitchell, A. G., etal J. Chem. Soc. Perkin Trans. 1,1992, 38: 2345). Thedisulfide-containing prodrugs, reported by Puech et al., Antiviral Res.,1993, 22: 155, can also be prepared from dichlorophosphonate and2-hydroxyethyl disulfide under the standard conditions.

Such reactive dichlorophosphonate intermediates can be prepared from thecorresponding phosphonic acids and the chlorinating agents e.g. thionylchloride (Starrett, et al, J. Med. Chem., 1994, 1857), oxalyl chloride(Stowell, et al, Tetrahedron Lett., 1990, 31: 3261), and phosphoruspentachloride (Quast, et al, Synthesis, 1974, 490). Alternatively, thesedichlorophosphonates can also be generated from disilyl phosphonateesters (Bhongle, et al, Synth. Commun., 1987, 17: 1071) and dialkylphosphonate esters (Still, et al, Tetrahedron Lett., 1983, 24: 4405;Patois, et al, Bull. Soc. Chim. Fr., 1993, 130: 485).

Furthermore, these prodrugs can be prepared from Mitsunobu reactions(Mitsunobu, Synthesis, 1981, 1; Campbell, J. Org. Chem., 1992, 52:6331), and other acid coupling reagents include, but not limited to,carbodiimides (Alexander, et al, Collect. Czech. Chem. Commun., 1994,59:1853; Casara, et al, Bioorg. Med. Chem. Lett., 1992, 2:145; Ohashi,et al, Tetrahedron Lett., 1988, 29: 1189), andbenzotriazolyloxytris-(dimethylamino)phosphonium salts (Campagne, et al,Tetrahedron Lett., 1993, 34: 6743). The prodrugs of formula 1 where R¹is the cyclic carbonate or lactone or phthalidyl can also be synthesizedby direct alkylation of free phosphonic acid with desired halides in thepresence of base such as NaH or diisopropylethylamine (Biller and MagninU.S. Pat. No. 5,157,027; Serafinowska et al. J. Med. Chem. 38:1372(1995); Starrett et al. J. Med. Chem. 37: 1857 (1994); Martin et al. J.Pharm. Sci. 76:180 (1987); Alexander et al., Collect. Czech. Chem.Commun, 59:1853 (1994); and EPO 0632048A1).

R¹ can also be introduced at an early stage of synthesis, when feasible.For example, compounds of formula 1 where R¹ is phenyl can be preparedby phosphorylation of 2-furanylpurines via strong base treatment (e.g.LDA) followed by chlorodiphenylphosphonate, as shown in the followingscheme. Alternatively, such compounds can be prepared by cyclization of5-diphenylphosphono-2-furaldehyde with 4,5-diaminopyrimidines asdescribed in section 5.

It is envisioned that compounds of formula 1 can be mixed phosphonateesters by combining the above described prodrugs (e.g. phenyl benzylphosphonate esters, phenyl acyloxyalkyl phosphonate esters, etc.). Forexample, the chemically combined phenyl-benzyl prodrugs are reported byMeier et al. Bioorg. Med. Chem. Lett., 1997, 7: 99.

The substituted cyclic propyl phosphonate esters of formula 5, can besynthesized by reaction of the corresponding dichlorophosphonate and thesubstituted 1,3-propanediol. The following are some of the methods toprepare the substituted 1,3-propanediols.

Synthesis of the 1,3-Propanediols Used in the Preparation of CertainProdrugs

The discussion of this step includes various synthetic methods for thepreparation of the following types of propane-1,3-diols: i)1-substituted; ii) 2-substituted; and iii) 1,2- or 1,3-annulated.Different groups on the prodrug part of the molecule i.e., on thepropanediol moiety can be introduced or modified either during thesynthesis of the diols or after the synthesis of the prodrugs.

i) 1-Substituted 1,3-Propanediols

Propane-1,3-diols can be synthesized by several well known methods inthe literature. Aryl Grignard additions to 1-hydroxypropan-3-al gives1-aryl-substituted propane-1,3-diols (path a). This method will enableconversion of various substituted aryl halides to1-arylsubstituted-1,3-propanediols (Coppi, et. al., J. Org. Chem., 1988,53, 911). Aryl halides can also be used to synthesize 1-substitutedpropanediols by Heck coupling of 1,3-diox-4-ene followed by reductionand hydrolysis (Sakamoto, et. al., Tetrahedron Lett., 1992, 33, 6845). Avariety of aromatic aldehydes can be converted to1-substituted-1,3-propanediols by vinyl Grignard addition followed byhydroboration (path b). Substituted aromatic aldehydes are also utilizedby lithium-t-butylacetate addition followed by ester reduction (path e)(Turner., J. Org. Chem., 1990, 55 4744). In another method, commerciallyavailable cinnamyl alcohols can be converted to epoxy alcohols undercatalytic asymmetric epoxidation conditions. These epoxy alcohols arereduced by Red-Al to result in enantiomerically pure propane-1,3-diols(path c). Alternatively, enantiomerically pure 1,3-diols can be obtainedby chiral borane reduction of hydroxyethyl aryl ketone derivatives(Ramachandran, et. al., Tetrahedron Lett., 1997, 38 761). Pyridyl,quinoline, and isoquinoline propan-3-ol derivatives can be oxygenated to1-substituted propan-1,3-diols by N-oxide formation followed byrearrangement under acetic anhydride conditions (path d) (Yamamoto, et.al., Tetrahedron, 1981, 37, 1871).

ii) 2-Substituted 1,3-Propanediols:

Various 2-substituted propane-1,3-diols can be made from commerciallyavailable 2-(hydroxymethyl)-1,3-propanediol. Triethylmethanetricarboxylate can be converted to the triol by completereduction or diol-monocarboxylic acid derivatives can be obtained bypartial hydrolysis and diester reduction (Larock, Comprehensive OrganicTransformations, VCH, New York, 1989). Nitrotriol is also known to givethe triol by reductive elimination (Latour, et. al., Synthesis, 1987, 8,742). The triol can be derivatized as a mono acetate or carbonate bytreatment with alkanoyl chloride, or alkylchloroformate, respectively(Greene and Wuts, Protective Groups in Organic Synthesis, John Wiley,New York, 1990). Aryl substitution can be made by oxidation to thealdehyde followed by aryl Grignard additions and the aldehyde can alsobe converted to substituted amines by reductive amination reactions.

iii) Annulated 1,3-Propanediols:

Prodrugs of formula 1 where V-Z or V-W are fused by three carbons aremade from cyclohexanediol derivatives. Commercially available cis,cis-1,3,5-cyclohexanetriol can be used for prodrug formation. Thiscyclohexanetriol can also be modified as described in the case of2-substituted propane-1,3-diols to give various analogues. Thesemodifications can either be made before or after formation of prodrugs.Various 1,3-cyclohexanediols can be made by Diels-Alder methodologyusing pyrone as the diene (Posner, et. al., Tetrahedron Lett., 1991, 32,5295). Cyclohexyl diol derivatives are also made by nitrile oxideolefin-additions (Curran, et. al., J. Am. Chem. Soc., 1985, 107, 6023).Alternatively, cyclohexyl precursors can be made from quinic acid (Rao,et. al., Tetrahedron Lett., 1991, 32, 547.)

(2) Deprotection of Phosphonate Esters

Compounds of formula 1 where R¹ is H may be prepared from phosphonateesters using known phosphate and phosphonate ester cleavage conditions.For example, alkyl phosphonate esters are generally cleaved by reactionwith silyl halides followed by hydrolysis of the intermediate silylphosphonate esters. Depending on the stability of the products, thesereactions are usually accomplished in the presence of acid scavengerssuch as 1,1,1,3,3,3-hexamethyldisilazane, 2,6-lutidine, etc. Varioussilyl halides can be used for this transformation, such aschlorotrimethylsilane (Rabinowitz J. Org. Chem., 1963, 28: 2975),bromotrimethylsilane (McKenna et al. Tetrahedron Lett., 1977, 155),iodotrimethylsilane (Blackburn et al. J. Chem. Soc., Chem. Commun.,1978, 870). Phosphonate esters can also be cleaved under strong acidconditions, such as hydrogen halides in acetic acid or water, and metalhalides (Moffatt et al. U.S. Pat. No. 3,524,846,1970). Phosphonateesters can also be converted to dichlorophosphonates with halogenatingagents (e.g. PCl₅, SOCl₂, BBr₃, etc. Pelchowicz et al. J. Chem. Soc.,1961, 238) and subsequently hydrolyzed to give phosphonic acids.Reductive reactions are useful in cleaving aryl and benzyl phosphonateesters. For example, aryl and benzyl phosphonate esters can be cleavedunder hydrogenolysis conditions (Lejczak et al. Synthesis, 1982, 412;Elliott et al. J. Med Chem., 1985, 28: 1208.) or dissolving metalreduction conditions (Shafer et al. J. Am. Chem. Soc., 1977, 99: 5118).(Elliott et al. J. Med. Chem., 1985, 28: 1208). Electrochemical (Shonoet al. J. Org. Chem., 1979, 44: 4508) and pyrolysis (Gupta et al. Synth.Commun., 1980, 10: 299) conditions have also been used to cleave variousphosphonate esters.

(3) Modification of C-8-substituted Purine Intermediates

8-Substituted purines are useful intermediates in the preparation ofcompounds of formula 1. 8-Halopurines, which are particularly usefulintermediates, are readily prepared using chemistry well described inthe literature. For example, N⁹-alkyladenines are halogenated at the C-8position using known halogenating agents (e.g. Br₂, NBS). 8-Alkylpurinecan be prepared through direct lithiation of purine followed by trappingwith electrophiles (e.g. alkyl halides, Barton et al. Tetrahedron Lett.,1979, 5877).

Functionaliztion of 8-halopurines can be accomplished under substitutionreaction conditions with nucleophiles such as amines, alcohols, azides,sulfides, and alkylthiols. It is advantageous to have the phosphonatemoiety as part of the nucleophiles. For example, substitution of8-bromopurine with aminoalkylphosphonates gives compounds of formula 1where X is alkylamino.

8-Halopurines can also be transformed into other 8-substituted purinesusing palladium catalyzed reactions (Heck Palladium Reagents in OrganicSynthesis; Academic Press: San Diego, 1985). For example, palladiumcatalyzed carbonylation reactions of 8-bromopurine in the presence ofalcohol gives 8-alkoxycarbonylpurines. Using known chemistry, the8-carboxylate group can be converted into other functional groups, suchas hydroxymethyl, halomethyl, formyl, carboxylic acid, carbamoyl, andthiocarbonyl groups. These functional groups are useful intermediatesfor the synthesis of compounds of formula 1. For example, 8-alkyl and8-arylpurines can be prepared from 8-halopurines via palladium catalyzedcoupling reactions with organotin (Moriarty et al. Tetrahedron Lett.,1990, 41: 5877), organoborane (Yatagai, Bull. Chem. Soc. Jpn., 1980, 53:1670), and other reagents known to couple with aryl halides. When thecoupling reagents contain the dialkylphosphonate group, the reaction isuseful for preparation of compounds of formula 5 where X is alkyl,alkenyl, alkynyl, and aryl. For example, 8-bromopurine can be coupledwith diethyl 1-tributylstannyl-3-allylphosphonate to give compounds offormula 5 where X is —CH═CHCH₂—. Subsequent hydrogenation reaction givescompounds of formula 5 where X is —CH₂CH₂CH₂—.

The phosphonate group can also be introduced by further modification ofthe 8-substituents. Substitutions of 8-haloalkyl or8-sulfonylalkylpurine with nucleophiles containing the phosphonate groupare useful for the preparation of compounds of formula 5 where X isalkylaminoalkyl, alkoxyalkyl, and alkylthioalkyl. For example, compoundsof formula 5 where X is —CH₂OCH₂— can be prepared from8-bromomethylpurine using hydroxymethylphosphonate esters and a suitablebase. It is possible to reverse the nature of the nucleophiles andelectrophiles for the substitution reactions, i.e. haloalkyl- and/orsulfonylalkylphosphonate esters can be substituted with purinescontaining a nucleophile at the C-8 position (such as 8-hydroxyalkyl,8-thioalkyl, and 8-aminoalkylpurines). For example, diethylphosphonomethyltriflate can be substituted by alcohols such as8-hydroxymethylpurine to give compounds of formula 5 where X is—CH₂OCH₂— (Phillion et al. Tetrahedron Lett. 1986, 27: 1477). Knownamide formation reactions are useful for the synthesis of compounds offormula 5 where X is alkylaminocarbonyl, alkoxycarbonyl,alkoxythiocarbonyl, and alkylthiocarbonyl. For example, coupling of8-purinecarboxylic acids with aminoalkylphosphonate esters givescompounds of formula 5 where X is alkylaminocarbonyl. For compounds offormula 5 where X is alkyl, the phosphonate group can also be introducedusing other common phosphonate formation methods, such asMichaelis-Arbuzov reaction (Bhattacharya et al. Chem. Rev., 1981, 81:415), Michaelis-Becker reaction (Blackburn et al. J. Organomet. Chem.,1988, 348: 55), addition reactions of phosphorus to electrophiles (suchas aldehydes, ketones, acyl halides, imines and other carbonylderivatives).

8-Azidopurines are useful for the preparation for compounds of formula 5where X is alkylamino and alkylcarbonylamino groups. For example,carboxylic acids (e.g. (RO)₂P(O)-alkyl-CO₂H) can be directly coupled to8-azidopurines to give 8-alkylcarbonylaminopurines (Urpi et al.Tetrahedron Lett., 1986, 27: 4623). Alternatively, 8-azidopurines canalso be converted to 8-aminopurines under reductive conditions, andsubsequently converted to 8-alkylaminocarbonyl- and 8-alkylaminopurinesusing known chemistry.

(4) Modification of Purines at Positions Other Than C-8

Compounds of formula 5 can be further modified to give intermediatesuseful for the synthesis of compounds of formula 1. For example,substitution reactions of 6-chloropurine by ammonia or alkylamines areuseful for the preparations of compounds of formula 5 where A is aminoand alkylamino groups.

E groups can be introduced by modifying existing 2-substituents ofpurine. For example, 2-halopurines, readily accessible from2-aminopurines via chemistry well described in the literature, can beconverted to other 2-substituted purines by, for example, nucleophilicsubstitution reactions; transition metal catalyzed reactions, etc. (J.Med. Chem., 1993, 36: 2938; Heterocycles, 1990, 30: 435).

E groups can also be introduced via metalation (e.g. lithiation, J. Org.Chem., 1997, 62(20), 6833) of the C-2-position and followed by additionof electrophiles which can be the desired E group or a substituent (e.g.tributylstannyl group) which can be converted to the desired E groupusing conventional chemistry.

It is envisioned that N⁹-substituted purines can be readily preparedfrom compounds of formula 5 where Y is H using, for example, standardalkylation reactions (with alkyl halide, or sulfonate), or Mitsunobureactions. Further elaborations of substituents on Y are also possible.

More importantly, combinatorial methods have been developed forsynthesis of 2- and N-9-substituted purines on solid-phase whichconceivably can be applied for the synthesis of purine FBPase inhibitors(Schultz, et al, Tetrahedron Lett., 1997, 38(7), 1161; J. Am. Chem.Soc., 1996, 118, 7430).

(5) Construction of the Purine Ring System

The purine ring system of compounds of formula 1 can be constructedusing 4,5-diaminopyrimidines and carboxylates or their derivatives (suchas aldehydes, amides, nitrites, ortho esters, imidates, etc.) (TownsendChemistry of Nucleosides and Nucleotides, Vol 1; Plenum Press, New Yorkand London, page 156-158). For example, alkyl and aryl aldehydes can becyclized with 4,5-diaminopyrimidines as shown below.

Intramolecular cyclization reactions of pyrimidine derivatives can alsobe used to construct the purine ring system. For example,5-acylamino-4-alkylaminopyrimidines are treated with phosphorusoxychloride and cyclized under basic conditions to give purinederivatives. This transformation can also be achieved using otherreagents (e.g. SiCl₄-Et₃N, Desaubry et al. Tetrahedron Lett., 1995, 36:4249). Imidazole derivatives are also useful for the construction ofpurine ring system via cyclization reactions to form the pyrimidine ring(Townsend Chemistry of Nucleosides and Nucleotides, Vol 1; Plenum Press,New York and London, page 148-156).

(6) Preparation of Diaminopyrimidine and Other Coupling Partners

Compounds of formula 4 are useful for the construction of purine ringsystems, and such compounds can be readily synthesized using knownchemistry. For example, the Y group can be introduced using anucleophilic substitution reaction involving an amine and4-halopyrimidines (Tetrahedron, 1984, 40: 1433). Alternatively,palladium catalyzed reactions (Wolfe et al. J. Am. Chem. Soc., 1996,118: 7215) can also be used. Reductive amination reactions (Synthesis,1975, 135) and alkylation with electrophiles (such as halides,sulfonates) are useful for the preparation of compounds of formula 4from 4-aminopyrimidines. The 5-amino group can be introduced using amineformation reactions such as nitration followed by reduction (Dhainant etal. J. Med. Chem., 1996, 39: 4099), arylazo compound formation followedby reduction (Lopez et al. Nucleosides & Nucleotides, 1996, 15: 1335),azide formation followed by reduction, or by rearrangement of carboxylicacid derivatives (e.g. Schmidt, Curtius, and Beckmann reactions).

Coupling of aromatic or aliphatic aldehydes, and carboxylic acidderivatives with attached phosphonate esters are particularly suited forthe preparation of compounds of formula 1 as described in section 5.Such phosphonate esters are prepared by lithiation of the aromatic ringusing methods well described in literature (Gschwend Org. React. 1979,26: 1) followed by addition of phosphorylating agents (e.g. ClPO₃R₂).Phosphonate esters can also be introduced by Arbuzov-Michaelis reaction(Brill Chem Rev., 1984, 84: 577) and transition metal catalyzed reactionwith alkyl halides and aryl halides or triflates (Balthazar et al. J.Org. Chem., 1980, 45: 5425; Petrakis et al. J. Am. Chem. Soc., 1987,109: 2831; Lu et al. Synthesis, 1987, 726). Alternatively, arylphosphonate esters can be prepared from aryl phosphates under anionicrearrangement conditions (Melvin Tetrahedron Lett., 1981, 22: 3375;Casteel et al. Synthesis, 1991, 691). Aryl phosphate esters can also beused to prepare compounds of Formula 1 where X is an oxyaryl group.N-Alkoxy aryl salts with alkali metal derivatives of dialkyl phosphonatecan be used to synthesize heteroaryl-2-phosphonate esters (Redmore J.Org. Chem., 1970, 35: 4114).

A second lithiation step can be used to incorporate the aldehydefunctionality, although other methods known to generate aromaticaldehydes can be envisioned as well (e.g. Vilsmeier-Hack reaction,Reimar-Teimann reaction etc.). In the second lithiation step, thelithiated aromatic ring is treated with reagents that either directlygenerate an aldehyde (e.g. DMF, HCO₂R, etc.) or with reagents that leadto a group that subsequently is transformed into an aldehyde group usingknown chemistry (e.g. alcohol, ester, cyano, alkene, etc.). It is alsoenvisioned that the sequence of these reactions can be reversed, i.e.the aldehyde moiety can be incorporated first followed by thephosphorylation reaction. The order of the reaction will depend on thereaction conditions and the protecting groups. Prior to thephosphorylation, it is also envisioned that it may be advantageous toprotect the aldehydes using a number of well-known steps (hemiacetal,hemiaminal, etc.,). The aldehyde is then unmasked after phosphorylation.(Protective groups in Organic Synthesis, Greene, T. W., 1991, Wiley, NewYork).

Formulations

Compounds of the invention are administered orally in a total daily doseof about 0.1 mg/kg/dose to about 100 mg/kg/dose, preferably from about0.3 mg/kg/dose to about 30 mg/kg/dose. The most preferred dose range isfrom 0.5 to 10 mg/kg (approximately 1 to 20 nmoles/kg/dose). The use oftime-release preparations to control the rate of release of the activeingredient may be preferred. The dose may be administered in as manydivided doses as is convenient. When other methods are used (e.g.intravenous administration), compounds are administered to the affectedtissue at a rate from 0.3 to 300 nmol/kg/min, preferably from 3 to 100nmoles/kg/min. Such rates are easily maintained when these compounds areintravenously administered as discussed below.

For the purposes of this invention, the compounds may be administered bya variety of means including orally, parenterally, by inhalation spray,topically, or rectally in formulations containing pharmaceuticallyacceptable carriers, adjuvants and vehicles. The term parenteral as usedhere includes subcutaneous, intravenous, intramuscular, andintraarterial injections with a variety of infusion techniques.Intraarterial and intravenous injection as used herein includesadministration through catheters. Oral administration is generallypreferred.

Pharmaceutical compositions containing the active ingredient may be inany form suitable for the intended method of administration. When usedfor oral use for example, tablets, troches, lozenges, aqueous or oilsuspensions, dispersible powders or granules, emulsions, hard or softcapsules, syrups or elixirs may be prepared. Compositions intended fororal use may be prepared according to any method known to the art forthe manufacture of pharmaceutical compositions and such compositions maycontain one or more agents including sweetening agents, flavoringagents, coloring agents and preserving agents, in order to provide apalatable preparation. Tablets containing the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipient which aresuitable for manufacture of tablets are acceptable. These excipients maybe, for example, inert diluents, such as calcium or sodium carbonate,lactose, calcium or sodium phosphate; granulating and disintegratingagents, such as maize starch, or alginic acid; binding agents, such asstarch, gelatin or acacia; and lubricating agents, such as magnesiumstearate, stearic acid or talc. Tablets may be uncoated or may be coatedby known techniques including microencapsulation to delay disintegrationand adsorption in the gastrointestinal tract and thereby provide asustained action over a longer period. For example, a time delaymaterial such as glyceryl monostearate or glyceryl distearate alone orwith a wax may be employed.

Formulations for oral use may be also presented as hard gelatin capsuleswhere the active ingredient is mixed with an inert solid diluent, forexample calcium phosphate or kaolin, or as soft gelatin capsules whereinthe active ingredient is mixed with water or an oil medium, such aspeanut oil, liquid paraffin or olive oil.

Aqueous suspensions of the invention contain the active materials inadmixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients include a suspending agent, such as sodiumcarboxymethylcellulose, methylcellulose, hydroxypropyl methylcelluose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia,and dispersing or wetting agents such as a naturally occurringphosphatide (e.g., lecithin), a condensation product of an alkyleneoxide with a fatty acid (e.g., polyoxyethylene stearate), a condensationproduct of ethylene oxide with a long chain aliphatic alcohol (e.g.,heptadecaethyleneoxycetanol), a condensation product of ethylene oxidewith a partial ester derived from a fatty acid and a hexitol anhydride(e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension mayalso contain one or more preservatives such as ethyl or n-propylp-hydroxy-benzoate, one or more coloring agents, one or more flavoringagents and one or more sweetening agents, such as sucrose or saccharin.

Oil suspensions may be formulated by suspending the active ingredient ina vegetable oil, such as arachis oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin. The oral suspensionsmay contain a thickening agent, such as beeswax, hard paraffin or cetylalcohol. Sweetening agents, such as those set forth above, and flavoringagents may be added to provide a palatable oral preparation. Thesecompositions may be preserved by the addition of an antioxidant such asascorbic acid.

Dispersible powders and granules of the invention suitable forpreparation of an aqueous suspension by the addition of water providethe active ingredient in admixture with a dispersing or wetting agent, asuspending agent, and one or more preservatives. Suitable dispersing orwetting agents and suspending agents are exemplified by those disclosedabove. Additional excipients, for example sweetening, flavoring andcoloring agents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachis oil, a mineral oil, such as liquid paraffin, ora mixture of these. Suitable emulsifying agents includenaturally-occurring gums, such as gum acacia and gum tragacanth,naturally occurring phosphatides, such as soybean lecithin, esters orpartial esters derived from fatty acids and hexitol anhydrides, such assorbitan monooleate, and condensation products of these partial esterswith ethylene oxide, such as polyoxyethylene sorbitan monooleate. Theemulsion may also contain sweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, such asglycerol, sorbitol or sucrose. Such formulations may also contain ademulcent, a preservative, a flavoring or a coloring agent.

The pharmaceutical compositions of the invention may be in the form of asterile injectable preparation, such as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according tothe known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butane-diol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils may conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid may likewise be used in the preparation ofinjectables.

The amount of active ingredient that may be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans maycontain 20 to 2000 μmol (approximately 10 to 1000 mg) of active materialcompounded with an appropriate and convenient amount of carrier materialwhich may vary from about 5 to about 95% of the total compositions. Itis preferred that the pharmaceutical composition be prepared whichprovides easily measurable amounts for administration. For example, anaqueous solution intended for intravenous infusion should contain fromabout 0.05 to about 50 μmol (approximately 0.025 to 25 mg) of the activeingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/hr can occur.

As noted above, formulations of the present invention suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion ora water-in-oil liquid emulsion. The active ingredient may also beadministered as a bolus, electuary or paste.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared bycompressing in a suitable machine the active ingredient in a freeflowing form such as a powder or granules, optionally mixed with abinder (e.g., povidone, gelatin, hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (e.g., sodiumstarch glycolate, cross-linked povidone, cross-linked sodiumcarboxymethyl cellulose) surface active or dispersing agent. Moldedtablets may be made by molding in a suitable machine a mixture of thepowdered compound moistened with an inert liquid diluent. The tabletsmay optionally be coated or scored and may be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropyl methylcellulose in varying proportionsto provide the desired release profile. Tablets may optionally beprovided with an enteric coating, to provide release in parts of the gutother than the stomach. This is particularly advantageous with thecompounds of formula 1 when such compounds are susceptible to acidhydrolysis.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored base, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert base such as gelatin and glycerin, or sucrose andacacia; and mouthwashes comprising the active ingredient in a suitableliquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

Formulations suitable for parenteral administration include aqueous andnon-aqueous isotonic sterile injection solutions which may containantioxidants, buffers, bacteriostats and solutes which render theformulation isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose sealed containers, for example, ampoules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carrier, for examplewater for injections, immediately prior to use. Injection solutions andsuspensions may be prepared from sterile powders, granules and tabletsof the kind previously described.

Preferred unit dosage formulations are those containing a daily dose orunit, daily sub-dose, or an appropriate fraction thereof, of a fructose1,6-bisphosphatase inhibitor compound.

It will be understood, however, that the specific dose level for anyparticular patient will depend on a variety of factors including theactivity of the specific compound employed; the age, body weight,general health, sex and diet of the individual being treated; the timeand route of administration; the rate of excretion; other drugs whichhave previously been administered; and the severity of the particulardisease undergoing therapy, as is well understood by those skilled inthe art.

Utility

FBPase inhibitors at the AMP site may be used to treat diabetesmellitus, lower blood glucose levels, and inhibit gluconeogenesis.

FBPase inhibitors at the AMP site may also be used to treat excessglycogen storage diseases. Excessive hepatic glycogen stores are foundin patients with some glycogen storage diseases. Since the indirectpathway contributes significantly to glycogen synthesis (Shulman, G. I.Phys. Rev. 1992. 72, 1019-1035), inhibition of the indirect pathway(gluconeogenesis flux) is expected to decrease glycogen overproduction.

FBPase inhibitors at the AMP site may also be used to treat or preventdiseases associated with increased insulin levels.

Increased insulin levels are associated with an increased risk ofcardiovascular complications and atherosclerosis (Folsom, et al.,Stroke, 1994, 25, 66-73; Howard, G. et al., Circulation 1996, 93,1809-1817). FBPase inhibitors are expected to decrease postprandialglucose levels by enhancing hepatic glucose uptake. This effect ispostulated to occur in individuals that are non-diabetic (orpre-diabetic, i.e. without elevated HGO or fasting blood glucoselevels). Increased hepatic glucose uptake will decrease insulinsecretion and thereby decrease the risk of diseases or complicationsthat arise from elevated insulin levels.

The compounds of this invention and their preparation can be understoodfurther by the examples which illustrate some of the processes by whichthese compounds are prepared. These examples should not, however, beconstrued as specifically limiting the invention and variations of theinvention, now known or later developed, are considered to fall withinthe scope of the present invention as hereinafter claimed.

EXAMPLES Example 1 Preparation of 5-diethylphosphono-2-furaldehyde (1)

Step A. A solution of 2-furaldehyde diethyl acetal (1 mmol) in THF wastreated with nBuLi (1 mmol) at −78° C. After 1 h, diethylchlorophosphate (1.2 mmol) was added and the reaction was stirred for 40min. Extraction and evaporation gave a brown oil.

Step B. The resulting brown oil was treated with 80% acetic acid at 90°C. for 4 h. Extraction and chromatography gave compound 1 as a clearyellow oil.

Alternatively this aldehyde can be prepared from furan as describedbelow.

Step C. A solution of furan (1 mmol) in diethyl ether was treated withTMEDA (N,N,N′N′-tetramethylethylenediamine) (1 mmol) and nBuLi (2 mmol)at −78° C. The solution was stirred for 0.5 h. at −78° C. and diethylchlorophosphate was added and stirred for another 1 h. Extraction anddistillation produced diethyl 2-furanphosphonate as a clear oil.

Step D. A solution of diethyl 2-furanphosphonate (1 mmol) in THF(tetrahydrofuran) was treated with LDA (1.12 mmol, lithiumN,N-diisopropylamide) at −78° C. for 20 min. Methyl formate (1.5 mmol)was added and the reaction was stirred for 1 h. Extraction andchromatography gave compound 1 as a clear yellow oil.

Preferably this aldehyde can be prepared from 2-furaldehyde as describedbelow.

Step E. A solution of 2-furaldehyde (1 mmol) and N,N′-dimethylethylenediamine (1 mmol) in toluene was refluxed with a Dean-Stark trap tocollect the resulting water. After 2 h the solvent was removed in vacuoand the residue was distilled to givefuran-2-(N,N′-dimethylimidazolidine) as a clear colorless oil, bp 59-61°C. (3 mm Hg).

Step F. A solution of furan-2-(N,N′-dimethylimidazolidine) (1 mmol) andTMEDA (1 mmol) in THF was treated with nBuLi (1.3 mmol) at −40 to −48°C. The reaction was stirred at 0° C. for 1.5 h and then cooled to −55°C. and treated with a solution of diethylchlorophosphate (1.1 mmol) inTHF. After stirring at 25° C. for 12 h the reaction mixture wasevaporated and subjected to extraction to give5-diethylphosphono-furan-2-(N,N′-dimethylimidazolidine) as a brown oil.

Step G. A solution of5-diethylphosphonofuran-2-(N,N′-dimethyl-imidazolidine) (1 mmol) inwater was treated with concentrated sulfuric acid until pH=1. Extractionand chromatography gave compound 1 as a clear yellow oil.

Example 2 Preparation ofN⁹-substituted-8-(2-(5-phosphono)furanyl)adenines

The preparation of N⁹-(2-phenethyl)-8-(2-(5-phosphono)furanyl)adenine isgiven as an example:

Step A. A solution of 5-amino-4,6-dichloropyrimidine (1 mmol) in nBuOHwas treated with Et₃N (1.2 mmol) and phenethylamine (1.05 mmol) at 80°C. After 12 h, the cooled reaction mixture was evaporated under vacuumand the residue was chromatographed to give6-chloro-5-amino-4-(phenethylamino)-pyrimidine as a yellow solid. mp156-157° C.; TLC: R_(f)=0.41, 50% EtOAc-hexane.

Step B. The 6-chloro-5-amino-4-(2-phenethylamino)pyrimidine (1 mmol) inDMSO was treated with 2-furaldehyde (1.5 mmol) and FeCl₃-silica (2.0mmol) at 80° C. After 12 h, the cooled reaction mixture was filtered andthe filtrate was evaporated under vacuum. Chromatography afforded6-chloro-N⁹-(2-phenethyl)-8-(2-furanyl)purine as a yellow solid. TLC:Rf=0.62, 50% EtOAc-hexane. Anal. calcd. for C₁₇H₁₃N₄OCl: C: 62.87; H:4.03; N: 17.25. Found: C: 62.66; H: 3.96; N: 17.07.

Step C. The 6-chloro-N⁹-(2-phenethyl)-8-(2-furanyl)purine (1 mmol) inTHF was treated with LDA (1.5 mmol) at −78° C. After 1 h, diethylchlorophosphate (5 mmol) was added and the reaction was stirred at −78°C. for 2 h and then quenched with saturated NH₄Cl. Extraction andchromatography gave6-chloro-N⁹-(2-phenethyl)-8-(2-(5-diethylphosphono)-furanyl)purine as ayellow solid. TLC: Rf=0.34,100% EtOAc.

Alternatively this type of compound can be prepared as follows:

Step D. A solution of 6-chloro-5-amino-4-(2-phenethylamino)pyrimidine (1mmol) in DMSO was treated with 5-diethylphosphono-2-furaldehyde (1, 1.5mmol), and FeCl₃-silica (2.0 mmol) at 80° C. After 12 h., the cooledreaction mixture was filtered and the filtrate was evaporated undervacuum. Chromatography afforded6-chloro-N⁹-(2-phenethyl)-8-(2-(5-diethyl-phosphono)furanyl)purine as ayellow solid. TLC: Rf=0.34, 100% EtOAc.

Step E.6-Chloro-N⁹-(2-phenethyl)-8-(2-(5-diethylphosphono)furanyl)-purine (1mmol) in THF-DMSO was treated with liquid ammonia (2 mL) in a steelbomb. After 12 h, the reaction was evaporated under vacuum and theresidue was purified through chromatography to giveN⁹-(2-phenethyl)-8-(2-(5-diethylphosphono)furanyl)adenine as a yellowsolid. TLC: Rf=0.12, 5% MeOH —CH₂Cl₂.

Step F. A solution ofN⁹-(2-phenethyl)-8-(2-(5-diethylphosphono)furanyl)-adenine (1 mmol) inacetonitrile was treated with bromotrimethylsilane (10 mmol). After 12h, the reaction was evaporated under vacuum and the residue was treatedwith a mixture of water and acetonitrile. The solid was collectedthrough filtration to giveN⁹-(2-phenethyl)-8-(2-(5-phosphono)furanyl)adenine (2.1). mp 242-2440°C.; Anal. calcd. for C₁₇H₁₆N₅O₄P+1.37H₂O: C: 50.16; 4.64; N: 17.21.Found: C: 48.95; H: 4.59; N: 16.80.

The following compounds were prepared according to above procedures:

2.2: N⁹-(2-cyclohexylethyl)-8-(2-(5-phosphono)furanyl)adenine. mp194-195° C.; Anal. calcd. for C₁₇H₂₂N₅O₄P+1 H₂O: C: 49.90; H: 5.90; N:17.10. Found: 50.20; H: 5.70; N: 17.10.

2.3: N⁹-(2-naphthylmethyl)-8-(2-(5-phosphono)furanyl)adenine. mp255-256° C.; Anal. calcd. for C₂₀H₁₆N₅O₄P+1 H₂O: C: 54.70; H: 4.10; N:15.90. 54.30; H: 4.20; N: 15.90.

2.4: N⁹-(1-(2,2-diphenyl)ethyl)-8-(2-(5-phosphono)furanyl)adenine. mp220-221° C.; Anal. calcd. for C₂₃H₂₀N₅O₄P+0.25 H20: C: 59.29; H: 4.43;N: 15.03. Found: C: 59.35; H: 4.25; N: 14.83.

2.5: N⁹-ethyl-8-(2-(5-phosphono)furanyl)adenine. mp >230° C.; Anal.calcd. for C₁₁H₁₂N₅O₄P+1 H₂O: C: 40.38; H: 4.31; N: 21.40. Found: C:40.45; H N: 21.44.

2.6: N⁹-isobutyl-8-(2-(5-phosphono)furanyl)adenine. mp >230° C.; Anal.calcd. for C₁₃H₁₆N₅O₄P: C: 46.30; H: 4.78; N: 20.76. Found: C: 46.00; H:4.61; N: 20.49.

2.7: N⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine. mp >230° C.; Anal.calcd. for C₁₄H₁₈N₅O₄P: C: 47.87; H: 5.16; N: 19.94. Found: C: 47.59; H:4.92; N: 19.53.

2.8: N⁹-adamentanemethyl-8-(2-(5-phosphono)furanyl)adenine. mp >250° C.;Anal. calcd. for C₂₀H₂₄N₅O₄P+0.5 H₂O+0.25 MeOH: C: 54.48; H: 5.87; N:15.69. Found: C: 54.62; H: 5.52; N: 15.36.

2.9: N⁹-cyclopropyl-8-(2-(5-phosphono)furanyl)adenine. mp >250° C.; MS(M+H) calcd for C₁₂H₁₃N₅O₄P: 322, found: 322.

2.10: N⁹-cyclopentyl-8-(2-(5-phosphono)furanyl)adenine. mp 220°C.(decomp); Anal. calcd. for C₁₄H₁₆N₅O₄P+1 H₂O: C: 45.78; H: 4.94; N:19.07. Found: C: 45.40; H: 4.79; N: 18.73.

2.11: N⁹-((2-ethoxy)phenyl)methyl-8-(2-(5-phosphono)furanyl)-adenine.mp >230° C.; Anal. calcd. for C₁₈H₁₈N₅O₅P+2 H₂O: C: 47.90; H: 4.91; N:C: 48.03; H: 4.53; N: 15.25.

2.12:N⁹-(1-(3-N,N-dimethylamino-2,2-dimethyl)propyl)-8-(2-(5-phosphono)-furanyl)adenine.mp >230° C.; Anal. calcd. for C₁₆H₂₃N₆O₄P+3 H₂O+0.5 HOAc+0.75 Na: C:41.19; H: 6.30; N: 16.95. Found: C: 41.35; H: 6.04; N: 16.57.

2.13:N⁹-(1-(3-hydroxyl-2,2-dimethyl)propyl)-8-(2-(5-phosphono)-furanyl)adenine.mp >230° C.; Anal. calcd. for C₁₄H₁₈N₅O₅P+0.25 H₂O: C: 45.23; H: 5.02;N: 18.83. Found: C: 45.40; H: 5.02; N: 18.44.

2.14:N⁹-(1-(3-chloro-2,2-dimethyl)propyl)-8-(2-(5-phosphono)-furanyl)adenine.mp >230° C.; Anal. calcd. for C₁₄H₁₇N₅O₄PCl+0.125 CHCl₃+0.06 AcOEt: C:42.50; H: 4.37; N: 17.25. Found: C: 42.62; H: 3.99; N: 16.87.

2.15: N⁹-(1-(3,3-dimethyl)butyl)-8-(2-(5-phosphono)furanyl)-adenine. mp230° C.; Anal. calcd. for C₁₅H₂₀N₅O₄P+1.25H₂O+0.13 AcOEt: C: 46.68; H:5.94; N: 17.56. Found: C: 46.67; H: 5.78; N: 17.35.

2.16:N⁹-(1,5,5-trimethyl-3-cyclohexenyl)methyl-8-(2-(5-phosphono)furanyl)-adenine.mp >230° C.; Anal. calcd. for C₁₉H₂₄N₅O₄P+0.5 H₂O+0.13 AcOEt: C: 53.54;H: 5.99; N: 16.01. Found: C: 53.67; H: 5.69; N: 15.75.

2.17: N⁹-(1-(1,2,2-trimethyl)propyl)-8-(2-(5-phosphono)-furanyl)adenine.mp >250° C.; Anal. calcd. for C₁₅H₂₀N₅O₄P+0.67 H₂O+0.13AcO Et: C: 47.74;N: 18.56. Found: C: 47.99; H: 5.39; N: 18.49.

2.18: 6-Amino-9-(3-(1-imidazolyl)propyl)-8-(2-(5-phosphono)furanyl)purine. mp 182-186° C.; Mass calcd.for C₁₅H₁₆N₇O₄P: 389. Found: M+H⁺=390.

Examples 3 Preparation ofN⁹-substituted-8-(2-phosphonoethylamino)adenines

Step A. Adenine (1 mmol) in DMF was treated with sodium hydride (1.2mmol) followed by benzyl bromide (1.2 mmol) at room temperature undernitrogen. The resulting mixture was warmed at 100° C. for 2 h. Thecooled reaction mixture was evaporated to dryness. Extraction andchromatography afforded N⁹-benzyladenine.

Step B. A solution of N⁹-benzyladenine (1 mmol) in acetic acid buffer(pH=4) was treated with bromine (1 mmol) at room temperature for 12 h.The reaction was quenched with 10% sodium sulfite solution and extractedwith dichloromethane. The combined extracts were dried (Na₂SO₄) andevaporated to dryness. Chromatography afforded N⁹-benzyl-8-bromoadenine.

Step C. A mixture of N⁹-benzyl-8-bromoadenine (1 mmol),aminoethylphosphonate (2 mmol), and sodium hydroxide (2 mmol) inethanol-water in a sealed tube was warmed at 110° C. under nitrogen.After 24 h the cooled reaction mixture was purified through preparativeHPLC to give N⁹-benzyl-8-(2-phosphonoethylamino)adenine (3.1). Exactmass calculated for C₁₄H₁₇N₆O₃P+H⁺: 349.1178. Found: 349.1180.

The following compounds were prepared according to this procedure:

3.2: N⁹-phenethyl-8-(2-phosphonoethylamino)adenine. mp 159-160° C.;Anal. calcd. for C₁₅H₁₉N₆O₃P+1.25 H₂O: C: 46.81; H: 5.63; N: 21.84.Found: C: 47.05; H: 5.63; N: 21.48.

3.3: N⁹-(2-naphthylmethyl)-8-(2-phosphonoethylamino)adenine. mp 189-190°C.; Anal. calcd. for C₁₈H₁₉N₆O₃P+1.5 H₂O: C: 50.82; H: 5.21; N: 19.76.Found: C: 50.71; H: 5.25; N: 19.54.

3.4: N⁹-cyclohexylethyl-8-(2-phosphonoethylamino)adenine. mp >250° C.;Anal. calcd. for C₁₅H₂₅N₆O₃P+0.33 H₂O: C: 48.12; H: 6.91; N: 22.44.Found: C: 48.12; H: 6.78; N: 22.15.

Example 4 Preparation of 8-(2-phosphonoethylamino)adenosines

A mixture of 8-bromoadenosine (1 mmol), aminoethylphosphonate (2 mmol),and sodium hydroxide (2 mmol) in ethanol-water in a sealed tube waswarmed at 110° C. under nitrogen. After 24 h the cooled reaction mixturewas purified through preparative HPLC to give8-(2-phosphonoethylamino)-adenosine (4.1). mp 175° C.; Anal. calcd. forC₁₂H₁₉N₆O₇P+0.5 H₂O: C: 36.10; H: 5.05; N: 21.05; P: 7.76. Found: C:36.08; H: 4.83; N: 20.36; P: 7.86.

The following compound was prepared in this manner:

4.2: 8-(2-Phosphonoethylamino)-5-deoxyadenosine as a white solid. mp220° C.; Anal. calcd. for C₁₂H₁₉N₆O₆P+1.5 H₂O: C: 35.92; H: 5.53; N:20.94. Found: C: 36.15; H: 5.12; N: 20.53.

Examples 5 Preparation of N⁹-alkyl-8-(phosphonomethoxymethyl)adenines

Step A. A mixture of N⁹-phenethyl-8-bromoadenine (1 mmol), tetrakis(triphenylphosphine)palladium (0.05 mmol), and triethylamine (5 mmol) inDMF in a sealed tube was warmed at 110° C. under 50 psi of carbonmonoxide. After 24 h the cooled reaction mixture was evaporated andpurified through chromatography to giveN⁹-phenethyl-8-methoxycarbonyladenine as a yellow solid. TLC: Rf=0.12,5% MeOH—CH₂Cl₂.

Step B. A solution of N⁹-phenethyl-8-methoxycarbonyladenine (1 mmol) intetrahydrofuran was treated with lithium aluminum hydride (1 mmol) at 0°C. for 1 h. Extraction and chromatography gaveN⁹-phenethyl-8-hydroxymethyl-adenine as a white solid. TLC: Rf=0.31, 10%MeOH—CH₂Cl₂.

Step C. A solution of N⁹-phenethyl-8-hydroxymethyladenine (1 mmol) inmethylene chloride was treated with phosphorus tribromide (1 mmol) at25° C. for 1 h. Extraction and chromatography gaveN⁹phenethyl-8-bromomethyl-adenine as a white solid. TLC: R_(f)=0.31,10%MeOH—CH₂Cl₂.

Step D. A solution of N⁹-phenethyl-8-bromomethyladenine (1 mmol) in DMFwas treated with a solution of diethyl hydroxymethylphosphonate sodiumsalt (1 mmol) in DMF at 25° C. for 1 h. Extraction and chromatographygave N⁹-phenethyl-8-diethylphosphonomethoxymethyladenine as a whitesolid. TLC: R_(f)=0.31,10% MeOH—CH₂Cl₂.

N⁹-phenethyl-8-diethylphosphonomethoxymethyladenine was subjected toStep F in Example 2 to giveN⁹-(2-phenethyl)-8-(phosphonomethoxymethyl)-adenine (5.1) as a whitesolid. mp >250° C.; Anal. calcd. for C₁₉H₂₂N₅O₄P+0.75 H₂O: C: 56.93; H:5.91; N: 10.48. Found: C: 56.97; H: 5.63; N: 10.28.

The following compounds were prepared according to this procedure:

5.2: N⁹-(2-cyclohexylethyl)-8-(phosphonomethoxymethyl)adenine. mp >250°C.; Anal. calcd. for C₁₅H₂₄N₅O₄P+1 H₂O: C: 46.51; H: 6.76; N: 18.08.Found: 46.47; H: 6.71; N: 17.91.

5.3: N⁹-(1-nanonyl)-8-(phosphonomethoxymethyl)adenine. mp 195-210° C.;Anal. calcd. for C₁₆H₂₈N₅O₄P+1 H₂O: C: 47.64; H: 7.50; N: 17.36. Found:C: 47.33; H: 7.34; N: 16.99.

5.4: N⁹-(3-cyclohexylpropyl)-8-(phosphonomethoxymethyl)adenine. mp230-250° C.; Anal. calcd. for C₁₉H₂₂N₅O₄P+0.9 H₂O+0.3 HBr: C: 45.34;16.52. Found: C: 45.74; H: 6.39; N: 16.18.

Alternatively this type of compound can also be prepared according tothe following procedure:

Step E. A solution of 6-chloro-5-amino-4-(neopentylamino)pyrimidine (1mmol) in diethyl ether was treated with pyridine (3 mmol), andacetoxyacetyl chloride (1.2 mmol) at 25° C. for 12 h. Extraction andchromatography afforded6-chloro-5-acetoxyacetyl-amino-4-neopentylaminopyrimidine as a yellowsolid. TLC: R_(f)=0.18, 30% EtOAc-hexane.

Step F. A solution of6-chloro-5-acetoxyacetylamino-4-neopentyl-aminopyrimidine (1 mmol) inphosphorus oxychloride was heated at reflux for 6 h. The cooled reactionmixture was evaporated to dryness and the residue was dissolved inpyridine and stirred at 25° C. for 20 h. Evaporation and chromatographyafforded 6-chloro-8-acetoxymethyl-N⁹-neopentylpurine as a yellow solid.TLC: R_(f)=0.51, 50% EtOAc-hexane.

Step G. A solution of 6-chloro-8-acetoxymethyl-N⁹-neopentylpurine (1mmol) in THF-water was treated with aqueous sodium hydroxide (1.5 mmol)at 0° C. for 0.5 h. Extraction and chromatography afforded6-chloro-8-hydroxymethyl-N⁹-neopentylpurine as a yellow gel. TLC:R_(f)=0.38, 33% EtOAc-hexane.

Step H. A solution of 6-chloro-8-hydroxymethyl-N⁹-neopentylpurine (1mmol) in methylene chloride was treated with phosphorus tribromide (1mmol) at 25° C. for 6 h. Extraction and chromatography afforded6-chloro-8-bromomethyl-N⁹-neopentylpurine as a white solid. TLC:R_(f)=0.64, 25% EtOAc-hexane.

Step I. A solution of 6-chloro-8-bromomethyl-N⁹-neopentylpurine (1 mmol)in DMF was treated with a solution of sodium diethylphosphono-methoxide(1 mmol) at 25° C. for 6 h. Extraction and chromatography afforded6-chloro-8-diethyl-phosphonomethoxymethyl-N⁹-neopentylpurine as a whitesolid. TLC: R_(f)=0.31, 50% EtOAc-hexane.

Step J. A solution of6-chloro-8-diethylphosphonomethoxymethyl-N⁹-neopentylpurine (1 mmol) inTHF-DMSO was treated with liquid ammonia (10 mmol) at 25° C. for 6 h.Extraction and chromatography afforded8-diethylphosphonomethoxymethyl-N⁹-neopentyladenine as a white solid.TLC: R_(f)=0.44,25% MeOH-EtOAc.

8-Diethylphosphonomethoxymethyl-N⁹-neopentyladenine was subjected toStep F in Example 2 to giveN⁹-neopentyl-8-(phosphonomethoxymethyl)-adenine (5.5) as a white solid.mp >250° C.; Anal. calcd. for C₁₂H₂₀N₅O₄P+1.5 H₂O: C: 40.45; H: 6.51; N:19.65. Found: C: 40.68; H: 6.35; N: 19.40.

Examples 6 Preparation ofN⁹-substituted-8-(1-(3-phosphono)propyl)adenines

Step A. A mixture of diethyl propargylphosphonate (1 mmol, preparedaccording to J. Org. Chem., 1993, 58(24), 6531.), tributyltin hydride(1.05 mmol), and AIBN (0.005 mmol) was heated at 60° C. for 18 h. Thecooled reaction mixture was purified through chromatography to givedimethyl (1-tributylstannyl)allyl-3-phosphonate as a yellow oil.

Step B. A solution of N⁹-(2-cyclohexylethyl)-8-bromoadenine (1 mmol),tetrakis(triphenylphosphine)palladium (0.1 mmol), and dimethyl(1-tributylstannyl)allyl-3-phosphonate (5 mmol) in DMF was warmed at 90°C. under nitrogen. After 2 h the cooled reaction mixture was evaporatedand purified through chromatography to giveN⁹-(2-cyclohexylethyl)-8-(3-dimethylphosphonopropene-1-yl)adenine as ayellow solid. TLC: R_(f)=0.48, 10% MeOH—CH₂Cl₂.

Step C. A solution ofN⁹-(2-cyclohexylethyl)-8-(3-dimethylphosphono-propene-1-yl)adenine inmethanol-acetic acid was stirred at room temperature under 50 psi of H₂for 12 h. Filtration and chromatography affordedN⁹-(2-cyclohexylethyl)-8-(1-(3-dimethylphosphono)propyl)adenine as ayellow solid. TLC: R_(f)=0.26,10% MeOH—CH₂Cl₂.

N⁹-(2-cyclohexylethyl)-8-(1-(3-dimethylphosphono)propyl)adenine wassubjected to Step F in Example 2 to giveN⁹-(2-cyclohexylethyl)-8-(1-(3-phosphono)propyl)adenine (6.1) as a whitesolid: mp 122-125° C.; Anal. calcd. for C₁₆H₂₆N₅O₃P+0.25 AcOH: C: 51.83;H: 7.12; N: 18.31. Found: C: 51.87; H: 6.96; N: 17.96.

6.2: N⁹-(2-phenethyl)-8-(1-(3-phosphono)propyl)adenine was also preparedin this manner as a solid. mp >250° C. Anal. calcd. for C₁₆H₂₀N₅O₃P+0.5H₂O: C: 51.89; H: 5.71; N: 18.91. Found: C: 51.81; H: 5.49; N: 18.66.

Examples 7 Preparation ofN⁹-(2-phenethyl)-8-(2-(5-phosphono)thienyl)adenine

Step A. A solution of 2-thienyllithium in THF (1 mmol) was added to asolution of diethyl chlorophosphate (1 mmol) at −78° C. under nitrogen.After 2 h the reaction was warmed to room temperature and quenched withbrine. Extraction and chromatography afforded2-diethylphosphonothiophene as a yellow oil. TLC: R_(f)=0.37, 50%EtOAc—hexane.

Step B. A solution of 2-diethylphosphonothiophene (1 mmol) in THF wastreated with nBuLi at −78° C. for 1 h. Tributyltin chloride was addedand stirred at −78° C. for 2 h and the reaction was quenched with waterand warmed to room temperature. Extraction and chromatography affordeddiethyl 2-(5-tributylstannyl)thienylphosphonate as a yellow oil. TLC:R_(f)=0.65, 50% EtOAc-hexane.

Step C. A mixture of N⁹-phenethyl-8-bromoadenine (1 mmol), tetrakis(triphenylphosphine)palladium (0.1 mmol), and diethyl2-(5-tributylstannyl)-thienylphosphonate (5 mmol) in DMF was warmed at80° C. under nitrogen. After 21 h the cooled reaction mixture wasevaporated to dryness. The dark oil was triturated with hexane and theresidue was dissolved in CH₂Cl₂ and filtered. The filtrate wasevaporated to giveN⁹-(2-phenethyl)-8-(2-(5-diethylphosphono)-thienyl)adenine as a yellowsolid. TLC: R_(f)=0.50, 10% MeOH—CH₂Cl₂.

N⁹-(2-phenethyl)-8-(2-(5-diethylphosphono)thienyl)adenine was subjectedto Step F in Example 2 to giveN⁹-(2-phenethyl)-8-(2-(5-phosphono)-thienyl)adenine (7.1) as a whitesolid. mp >250° C.; Anal. calcd. for C₁₇H₁₆N₅O₃SP+0.5H₂O: C: 49.76; H:4.17; N: 17.07. Found: C: 50.07; H: N: 17.45.

N⁹-(2-phenethyl)-8-(2-(5-phosphono)thienyl)adenine can also be made viaa cyclization reaction between5-diethylphosphono-2-thiophene-carboxaldehyde (prepared from2-thienyllithium as described in Steps C and D of Example 1) asdescribed in Example 2.

Example 8 Preparation ofN⁹-(2-cyclohexylethyl)-8-(phosphonomethylthio)-adenine

Step A. A mixture of N⁹-(2-cyclohexylethyl)-8-bromoadenine (1 mmol), andK₂S (4 mmol) in ethanol was warmed at 110° C. for 7 h, and at 85° C. for12 h. The cooled reaction mixture was filtered, evaporated and purifiedthrough chromatography to giveN⁹-(2-cyclohexylethyl)-8-thiohydroxyadenine as a yellow solid. TLC:R_(f)=0.26, 5% MeOH—CH₂Cl₂

Step B. A mixture of N⁹-(2-cyclohexylethyl)-8-thiohydroxyadenine (1mmol), K₂CO₃ (4 mmol), and diethyl chloromethylphosphonate (3 mmol) inDMF was stirred at room temperature for 48 h. Extraction andchromatography gaveN⁹-(2-cyclohexylethyl)-8-diethylphosphono-methylthioadenine. TLC:R_(f)=0.35, 10% MeOH—EtOAc.

N⁹-(2-Cyclohexylethyl)-8-diethylphosphonomethylthioadenine was subjectedto Step F in Example 2 to giveN⁹-(2-cyclohexylethyl)-8-(phosphonomethylthio)adenine (8.1) as a whitesolid. mp 240-243° C.; Anal. Calcd. for C₁₄H₂₂N₅O₃SP+1.25H₂O: C: 42.69;H: 5.95; N: 17.54. Found: C: 42.62; H: 6.03; N: 17.80.

Example 9 Preparation of6-chloro-9-phenethyl-8-(2-(5-phosphono)furanyl)purine

6-Chloro-N⁹-phenethyl-8-(2-(5-diehtylphosphono)furanyl)purine (Step C inExample 2) was subjected to procedure of Step F in Example 2 to givecompound 9.1 as a yellow solid. mp >200° C.; Anal. calcd. forC₁₇H₁₄N₄O₄PCl+2 H₂O+0.28 HBr: C: 44.06; H: 3.98; N: 12.09. Found: C:43.86; H: 3.59; N: 12.02.

Example 10 Preparation ofN⁶,N⁹-substituted-8-(2-(5-phosphono)furanyl)adenines

A solution of6-chloro-N⁹-substituted-8-(2-(5-diethylphosphono)furanyl)-purine (1mmol) in DMSO was treated with alkylamine at 100° C. for 12 h.Evaporation and chromatography gaveN⁶,N⁹-substituted-8-(2-(5-diethyl-phosphono)furanyl)adenines.

The title compounds were obtained by subjectingN⁶,N⁹-substituted-8-(2-(5-diethylphosphono)furanyl)adenines to theprocedure of Step F in Example 2.

The following compounds were prepared in this manner:

10.1: 6-Dimethylamino-N⁹-phenethyl-8-(2-(5-phosphono)furanyl)purine as awhite solid. mp >200° C.; Anal. calcd. for C₁₉H₂₀N₅O₄P: C: 55.2; H: 4.8;N: 16.9. Found: C: 54.9; H: 4.9; N: 16.6.

10.2: 6-Methylamino-N⁹-phenethyl-8-(2-(5-phosphono)furanyl)purine as awhite solid. mp 242° C.; Anal. calcd. for C₁₈H₁₈N₅O₄P+1 H₂O: C: 51.8; H:4.8; N: 16.8. Found: C: 51.7; H: 4.8; N: 16.7.

Example 11 Preparation of2-methylthio-6-amino-N-isobutyl-8-(2-(5-phosphono)furanyl)purine and2-methylsulfonyl-6-amino-N⁹-isobutyl-8-(2-(5-phosphono)furanyl)purine

Step A: 2-Methylthio-4,5,6-triaminopyrimidine and5-diethylphosphono-2-furaldehyde was subjected to the procedures of StepD in Example 2 to give6-amino-2-methylthio-8-(2-(5-diethylphosphono)furanyl)purine as a yellowsolid. TLC: R_(f)=0.27, 80% EtOAc—hexane.

Step B: 6-Amino-2-methylthio-8-(2-(5-diethylphosphono)furanyl)purine wasalkylated with isobutyl bromide following the procedures of Step A inExample 3 to give6-amino-N⁹-isobutyl-2-methylthio-8-(2-(5-diethylphosphono)-furanyl)purineas a yellow solid. TLC: R_(f)=0.27, 80% EtOAc—hexane.

Step C:6-Amino-N⁹-isobutyl-2-methylthio-8-(2-(5-diethyl-phosphono)-furanyl)purinewas subjected to Step F in Example 2 to give6-amino-N⁹-isobutyl-2-methylthio-8-(2-(5-phosphono)-furanyl)purine(11.1) as a white solid. mp 220° C.; Anal. calcd. for C₁₄H₁₈N₅O₄PS+0.25HBr+0.25 EtOAc: C: 42.33; H: 4.8; N: 16.45. Found: C: 42.42; H: 4.53; N:16.39.

Step D: A solution of6-amino-N⁹-isobutyl-2-methylthio-8-(2-(5-diethyl-phosphono)furanyl)purine(1 mmol) in 50 mL of methanol was cooled to 0° C. and treated with anacetone solution of Oxone (1.6 mmol). After stirring for 3 h at 25° C.the reaction was extracted and then chromatographed to give6-amino-N⁹-isobutyl-2-methylsulfonyl-8-(2-(5-diethylphosphono)furanyl)purineas a white solid. TLC: R_(f)=0.24,100% EtOAc.

Step E:6-Amino-N⁹-isobutyl-2-methylsulfonyl-8-(2-(5-diethylphosphono)-furanyl)purinewas subjected to Step F in Example 2 to give6-amino-N⁹-isobutyl-2-methylsulfonyl-8-(2-(5-phosphono)furanyl)purine(11.2) as a white solid. mp 240° C. (decomp); Anal. calcd. forC₁₄H₁₈N₅O₆PS+0.5 H₂O: C: 39.62; H: 4.51; N: 16.5. Found: C: 39.77; H:4.44; N: 16.12

Example 12 Preparation of6-amino-N⁹-neopentyl-8-(2-(3.4-dichloro-5-phosphono)furanyl)purine

Step A: A solution of 3,4-dichloro-2-furoic acid (1 mmol) in diethylether was treated with LDA (3 mmol) at −78° C. for 30 min and thentreated with diethyl chlorophosphate (3.5 mmol) at −78° C. for 1 h. Thereaction was quenched and extracted to give5-diethylphosphono-3,4-dichloro-2-furoic acid as a yellow foam.

Step B: A solution of 5-diethylphosphono-3,4-dichloro-2-furoic acid (1mmol) in methylene chloride was treated with oxalyl chloride and DMF at25° C. for 1 h. The reaction mixture was evaporated and the residue wasdissolved in diethyl ether and treated with a solution of4-chloro-5-amino-6-neopentyl-aminopyrimidine (1 mmol) and pyridine (3mmol) in diethyl ether at 25° C. for 16 h. Extraction and chromatographygave4-chloro-5-(2-(3,4-dichloro-5-diethylphosphono)furoyl)amino-6-neopentylaminopyrimidineas a yellow solid. TLC: R_(f)=0.4, 50% EtOAc-hexane.

Step C: A solution of4-chloro-5-(2-(3,4-dichloro-5-diethylphosphono)-furoyl)amino-6-neopentylaminopyrimidine(1 mmol) in dichloromethane was treated with silicone tetrachloride (2.5mmol) and triethylamine (2.5 mmol) at 45° C. for 18 h. The cooledreaction mixture was subjected to extraction and chromatography to give6-chloro-N⁹-neopentyl-8-(2-(3,4-dichloro-5-diethyl-phosphono)furanyl)purineas a yellow solid. TLC: R_(f)=0.28, 50% EtOAc-hexane.

Step D:6-Chloro-N⁹-neopentyl-8-(2-(3,4-dichloro-5-diethylphosphono)-furanyl)purinewas subjected to Steps E and F in Example 2 to give6-amino-N⁹-neopentyl-8-(2-(3,4-dichloro-5-phosphono)furanyl)purine(12.1) as a white solid. mp >250° C.; Anal. calcd. forC₁₄H₁₆N₅O₄PCl₂+0.5 H₂O+0.15 EtOAc: C: 39.64; H: 4.15; N: 15.83. Found:C: 39.82; H: 3.88; N: 15.46.

Example 13 Preparation of hydroxyethyldisulfidylethylphosphonate diester

A suspension of 8-(2-(5-phosphono)furanyl)-N⁹-phenethyladenine (1 mmol)in thionyl chloride (5 mL) was warmed at reflux for 4 h. The cooledreaction mixture was evaporated to dryness and the resulting yellowresidue was treated with a solution of 2-hydroxyethyl disulfide (4mmol), pyridine (2.5 mmol) in methylene chloride. After stirring at 25°C. for 4 h the reaction was subjected to extraction and chromatographyto give two compounds:

13.1:N⁹-phenethyl-8-(bis(6′-hydroxy-3′,4′-disulfide)hexylphosphono)furanyl)-adenine.Anal. calcd for C₂₅H₃₂N₅O₆S₄P+0.5 DMSO+1.5 H₂O: C: 43.15; H: 5.29; N:9.68. Found: C: 43.38; H: 4.93; N: 9.34.

13.2:N⁹-phenethyl-8-((3′,4′-disulfide)nonacyclicphosphono)furanyladenine.Anal. calcd for C₂₁H₂₂N₅O₄S₂P+DMSO: C: 47.49; H: 4.85; N: 12.04. Found:C: 47.93; H: 4.60; N: 11.76.

Example 14 Preparation of substituted benzyl phosphonate diesters

A suspension of 8-(2-(5-phosphono)furanyl)-N⁹-phenethyladenine (1 mmol)in thionyl chloride (5 mL) was warmed at refluxing 4 h. The cooledreaction mixture was evaporated to dryness and a solution of theresulting yellow residue was added to a solution of the correspondingbenzyl alcohol (4 mmol), and pyridine (2.5 mmol) in methylene chloride.After stirring at 25° C. for 4 h the reaction mixture was subjected toextraction and chromatography to give the title compounds.

14.1:N⁹-phenethyl-8-(2-(5-(bis-(3-bromo-4-methoxy)benzyl)phosphono)-furanyl)adenine.Molecular mass calculated for C₃₃H₃₀N₅O₆Br₂P+H⁺: 784. Found: 784.

14.2:N⁹-phenethyl-8-(2-(5-(bis-(3-cyano-4-methoxy)benzyl)phosphono)-furanyl)adenine.Anal. calcd. for C₃₅H₃₀N₇O₆P+0.5 H₂O: C: 61.40; H: 4.56; N: 14.32.Found: C: 61.45; H: 4.51; N: 14.18.

14.3:N⁹-neopentyl-8-(2-(5-(bis-(4-acetoxy)benzyl)phosphono)furanyl)adenine.Anal. calcd. for C₃₂H₃₄N₅O₈P+0.6 H₂O: C: 58.37; H: 5.39; N: 10.64.Found: C: 58.11; H: 5.28; N: 10.42.

N⁹-neopentyl-8-(2-(5-(bis-(3-phthalidyl-2-ethyl)phosphono)furanyl)-adenineis also prepared following the above described procedure using2-(3-phthalidyl)ethanol which was prepared from phthalide-3-acetic acidin Example 27.

This reaction procedure can also be used to prepare diaryl esterprodrugs of phosphonates, such as substituted phenyl esters ofphosphonate.

Example 15 Preparation of6-amino-8-(2-(5-diphenylphosphono)furanyl)-N⁹-(2-phenyl)ethylpurine

Step A. A suspension of 6-chloro-8-(2-furanyl)-N⁹-phenethylpurine (1mmol) in THF at −78° C. was treated with LDA (1.3 mmol) for 1 h. Then asolution of diphenyl chlorophosphate in THF was added and the reactionwas stirred at −78° C. for another hour. The reaction was warmed to 0°C. and quenched with aqueous saturated sodium bicarbonate. Extractionand chromatography gave6-chloro-8-(2-(5-diphenylphosphono)furanyl)-N⁹-phenethylpurine as awhite solid. mp 117-118° C.

Step B. A solution of6-chloro-8-(2-(5-diphenylphosphono)furanyl)-N⁹-phenethylpurine (1 mmol)in DMF was treated with sodium azide (4 mmol) and triphenylphosphine (4mmol) at room temperature for 3 h. Filtration, evaporation of thefiltrate followed by chromatography gave6-triphenyl-phosphonoimino-8-(2-(5-diphenylphosphono)furanyl)-N⁹-(2-phenyl)ethylpurineas a beige foam.

Step C. A solution of6-triphenylphosphonoimino-8-(2-(5-diphenyl-phosphono)furanyl)-9-(2-phenyl)ethylpurine(1 mmol) in THF was treated with aqueous hydrogen chloride at roomtemperature for 24 h. Evaporation and chromatography gave6-amino-8-(2-(5-diphenylphosphono)furanyl)-N⁹-(2-phenyl)ethylpurine(15.1) as a pale yellow solid. mp 196-197° C.; Anal. calcd. forC₂₉H₂₄N₅O₄P: C: 64.80; H: 4.50; N: 13.03; P: 5.76. Found: C: 64.50; H:4.47; N: 12.98; P: 5.46.

Example 16 Preparation of acyloxymethylphosphonate diesters

A solution of 8-(2-(5-phosphono)furanyl)-N⁹-phenethyladenine (1 mmol) inacetonitrile and N,N,N-diisopropylethylamine (5 mmol) was treated withacyloxymethyl iodide (4 mmol) at 0° C. for 24 h. Extraction andchromatography gave the title compounds.

The following compounds were prepared according to this procedure:

16.1:6-Amino-9-phenethyl-8-(2-(5-diisobutyrylmethylphosphono)furanyl)-purine.Anal. calcd for C₂₇H₃₂N₅O₈P: C: 55.40; H: 5.50; N: 12.00. Found: C:55.60; H: 5.60; N: 11.80.

16.2:6-Amino-9-(2-cyclohexylethyl)-8-(2-(5-diisobutyrylmethylphosphono)-furanyl)purine.Anal. calcd for C₂₇H₃₈N₅O₈P+0.7 H₂O: C: 53.70; H: 6.60; N: 11.60. Found:C: 54.00; H: 6.50; N: 11.20.

16.3:6-Amino-9-ethyl-8-(2-(5-diisobutyrylmethylphosphono)-furanyl)purine.Anal. calcd for C₂₁H₂₈N₅O₈P: C: 49.51; H: 5.54; N: 13.75. Found: C:49.75; H: 5.37; N: 13.76.

16.4:6-Amino-9-neopentyl-8-(2-(5-diisobutyrylmethylphosphono)furanyl)-purine.Anal. calcd for C₂₄H₃₄N₅O₈P: C: 52.27; H: 6.21; N: 12.70. Found: C:52.40; H: 6.27; N: 12.41.

16.5:6-Amino-9-neopentyl-8-(2-(5-dipivaloxymethylphosphono)furanyl)purine.Anal. calcd for C₂₆H₃₈N₅O₈P+0.2 EtOAc: C: 53.90; H: 6.68; N: 11.73.Found: C: 54.10; H: 6.80; N: 11.42.

6-Amino-9-phenethyl-8-(2-(5-bis-(3-(5,6,7-trimethoxy)phthalidyl)-phosphono)furanyl)purine(16.6) was also synthesized following this procedure using3-bromo-5,6,7-trimethoxyphthalide as the alkylating reagent to give thetitled compound as a white solid after preparative HPLC purification. mp155-160° C.; Anal. calcd. for C₃₉H₃₆N₅O₁₄P+H₂O: C: 55.26; H: 4.52;Found: C: 54.89; H: 4.75; N: 8.21.

Example 17 Preparation of 5-methyl-4-hydroxymethyl-2-oxo-1,3-dioxolene

A solution of 4,5-dimethyl-2-oxo-1,3-dioxolene (1 mmol) and seleniumdioxide (2.5 mmol) in dioxane was heated at reflux for 1 h. Evaporation,extraction and chromatography gave5-methyl-4-hydroxymethyl-2-oxo-1,3-dioxolene as a yellow oil. TLC:R_(f)=0.5, 5% MeOH-dichloromethane.

Example 18 Preparation of (5-substituted 2-oxo-1,3-dioxolen-4-yl)methylphosphonate prodrugs

A solution of N⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine (1 mmol) inDMF and 2 mmol of sodium hydride is treated with5-methyl-4-bromomethyl-2-oxo-1,3-dioxolene (4 mmol, prepared accordingto Chem. Pharm. Bull. 1984, 32(6), 2241) at 25° C. for 24 h. Extractionand chromatography givesN⁹-neopentyl-8-(2-(5-bis(5-methyl-2-oxo-1,3-dioxolen-4-yl)methylphosphono)-furanyl)adenine.

Alternatively,N⁹-neopentyl-8-(2-(5-bis(5-methyl-2-oxo-1,3-dioxolen-4-yl)methylphosphono)-furanyl)adenine is prepared fromN⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine and5-methyl-4-hydroxymethyl-2-oxo-1,3-dioxolene (prepared from4,5-dimethyl-2-oxo-1,3-dioxolene as described in Example 17) accordingto procedures of Example 14.

Example 19 Preparation of 2-(6-amino-N⁹-neopentylpurin-8-yl)phenylphosphonate

Step A. Triethylamine (1.1 mmol) was added slowly to an ice-cooledsolution of 6-chloro-N⁹-neopentyl-8-(2-hydroxyphenyl)purine (1 mmol) anddiethyl phosphite (1 mmol) in carbontetrachloride. The reaction wasstirred at room temperature overnight. Triethylamine hydrochloride wasprecipitated as a white solid mass. Extraction and chromatography gavediethyl 2-(6-Chloro-N⁹-neopentylpurin-8-yl)phenyl phosphate.

Step B. Diethyl 2-(6-Chloro-N⁹-neopentylpurin-8-yl)phenyl phosphate wassubjected to Step E and F in Example 2 to give the title compound(19.1). mp >250° C.; Anal. calcd. for C₁₆H₂₀N₅O₄P+1.25 H₂O: C: 48.06; H:5.67; N: 17.51. Found: C:48.42; H: 5.42; N: 17.15.

Example 20 Preparation ofN⁹-neopentyl-8-(1-(2-phosphono)imidazolemethyl)adenine

Step A. A solution of 1-benzylimidazole (1.1 mmol) in THF was treatedwith LDA (1.1 mmol) at −78° C. for 1 h, and followed by addition ofdiethyl chlorophosphate (2 mmol), and stirred for 2 h. Extraction andchromatography gave 1-benzyl-2-diethylphosphonoimidazole as a yellowoil. TLC: R_(f)=0.35, 80% EtOAc-hexane.

Step B. A solution of 1-benzyl-2-diethylphosphonoimidazole (1 mmol) inEtOH was treated with palladium on carbon (10%) at 25° C. under 1atmosphere of hydrogen for 19 h. Filtration and evaporation gave2-diethyl-phosphonoimidazole as a white solid. TLC: R_(f)=0.05, 80%EtOAc-hexane.

Step C. A solution of 8-bromomethyl-6-chloro-N⁹-neopentylpurine (1 mmol,Step H of Example 5), 2-diethylphosphonoimidazole (2.5 mmol), andN,N,N-diisopropylethylamine (2.5 mmol) in acetonitrile was stirred at25° C. for 48 h. Extraction and chromatography gave6-chloro-N⁹-neopentyl-8-(1-(2-diethylphosphono)imidazolemethyl)purine.

Step D.6-Chloro-N⁹-neopentyl-8-(1-(2-diethylphosphono)imidazole-methyl)purinewas subjected to Steps E and F in Example 2 to give the title compound(20.1). mp >250° C.; MS (M+H) calcd. for C₁₄H₂₀N₇O₃P: 366; found: 366.

Example 21 Preparation ofN⁹-phenethyl-8-(phosphonomethylaminocarbonyl)adenine

Step A. N⁹-phenethyl-8-(methoxycarbonyl)adenine (1 mmol, prepared as inStep A of Example 5) was treated with sodium hydroxide (1.2 mmol) inTHF:MeOH:H₂O (3:2:1) at 25° C. for 1.5 h. The reaction mixture wasevaporated to dryness, and the residue was dissolved in DMF, treatedwith diethyl aminomethylphosphonate (1.5 mmol), EDCl(1-(3-dimethyl-aminopropyl)-3-ethylcarbodiimide hydrochloride, 1.3mmol), HOBt (1-hydroxy-benzotriazole hydrate, 1.5 mmol), and stirred at25° C. for 24 h. Extraction and chromatography gaveN⁹-phenethyl-8-(diethylphosphonomethyl-aminocarbonyl)adenine as a whitesolid. TLC: R_(f)=0.1, EtOAc.

Step B. N⁹-phenethyl-8-(diethylphosphonomethylaminocarbonyl)adenine (1mmol) was subjected to Step F in Example 2 to give the title compound(21.1). mp >250° C.; Anal. calcd. for C₁₅H₁₇N₆O₄P+0.17 Toluene: C:47.32; H: 4.50; N: 20.45. Found: C:47.67; H: 4.57; N: 20.78.

Example 22 Preparation of 2-substitutedN⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine

Step A. A solution of 2-amino-4,6-dichloropyremidine (1 mmol),neopentylamine (1.05 mmol), and triethylamine (2 mmol) in n-butanol wasstirred at 110° C. for 12 h. Extraction and chromatography gave2-amino-4-chloro-6-neopentylpyrimidine as a yellow solid. TLC:R_(f)=0.2, 30% EtOAc-hexane.

Step B. A mixture of 2-amino-4-chloro-6-neopentylpyrimidine (1 mmol),sodium acetate (14 mmol), acetic acid (86 mmol), and4-chlorobenzene-diazonium hexafluorophosphate (1.15 mmol) in water wasstirred at 25° C. for 12 h. Extraction and evaporation gave a yellowsolid which was treated with zinc dust (10 mmol) and acetic acid (5.54mmol) in EtOH-H₂O at 80° C. for 1 h. Extraction and chromatography gave4-chloro-2,5-diamino-6-neopentyl-pyrimidine as a yellow solid. TLC:R_(f)=0.25, 50% EtOAc-hexane.

Step C. 4-Chloro-2,5-diamino-6-neopentylpyrimidine was subjected to StepD, E, F in Example 2 to give2,6-diamino-N⁹-neopentyl-8-(2-(5-phosphono)furanyl)purine (22.1) as ayellow solid. mp 240° C. (decomp); Anal. calcd. for C₁₄H₁₉N₆O₄P+2.2HBr+0.5 acetone: C: 32.47; H: 4.25; N: 14.66. Found: C:32.31; H: 4.51;N: 14.85.

Similarly, 2-methylthio-N⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine

(22.2) was also prepared from 4-amino-6-chloro-2-methylthiopyrimidine asa yellow solid. mp >250; Anal. calcd. for C₁₅H₂₀N₅O₄PS+0.2 CH₂Cl₂+0.1toluene: C: 45.08; H: 5.04; N: 16.53. Found: C:45.27; H: 5.34; N: 16.24.

Example 23 Preparation of alkyloxycarbonyloxyalkyl phosphonate esters

A solution of N⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine (1 mmol) in5 mL of anhydrous DMF is treated withN,N′-dicyclohexyl-4-morpholinecarboxamidine (5 mmol), andisopropyloxycarbonyloxymethyl iodide (5 mmol) which is prepared from thecommercially available chloromethyl chloroformate according to thereported procedure, Nishimura et al. J. Antibiotics, 1987, 40(1), 81-90.The reaction mixture is stirred for 24 h at room temperature and thesolvent is removed under reduced pressure. The resulting syrup ischromatographed on silica with 50%/50% EtOAc/Hexane to yieldN⁹-neopentyl-8-(2-(5-diisopropyloxycarbonyloxymethyl phosphono)furanyl)adenine.

Other alkyloxycarbonyloxymethyl, aryloxycarbonyloxymethyl , alkyl- andarylthiocarbonyloxymethyl phosphonate esters can also be preparedfollowing the above described procedure.

Example 24 Preparation of 1-substituted-1,3-propanediol cyclic esters ofpurine phosphonates

Step A(J. Org. Chem. 1 957. 22 589)

To a solution of 2-pyridine propanol (72.9 mmol) in acetic acid (75 mL)was added 30% hydrogen peroxide slowly. The reaction mixture was heatedto 80° C. for 16 h. The reaction was concentrated under vacuum and theresidue was dissolved in acetic anhydride (100 mL) and heated at 110° C.overnight. Acetic anhydride was evaporated upon completion of reaction.Chromatography of the mixture by eluting with methanol-methylenechloride (1:9) resulted in 10.5 g of pure2-(1-(1,3-diacetoxy)propyl)pyridine.

Step B. To a solution of 2-(1-(1,3-diacetoxy)propyl)pyridine (21.1 mmol)in methanol-water (3:1, 40 mL) was added potassium carbonate (105.5mmol). After stirring for 3 h at room temperature, the reaction mixturewas concentrated. The residue was chromatographed by eluting withmethanol-methylene chloride (1:9) to give 2.2 g of crystalline2-(1-(1,3-dihydroxy)propyl)pyridine.

Step C. A suspension of N⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine(1 mmol) in 5 mL of thionyl chloride was heated at reflux temperaturefor 4 h. The reaction mixture was cooled and evaporated to dryness. Tothe resulting residue was added a solution of2-(1-(1,3-dihydroxy)propyl)pyridine (1 mmol) and pyridine (2.5 mmol) in3 mL of methylene chloride. After stirring at 25° C. for 4 h thereaction was subjected to work up and chromatography to giveN⁹-neopentyl-8-(2-(5-(1-(2-pyridyl)propan-1,3-yl)phosphono)furanyl)adenine(24.1) as a sticky solid. Anal. Calcd. for C₂₂H₂₅N₆O₄P+0.75 H₂O+1.0 HCl:C:50.97; H: 5.35; N: 16.21. Found: C:51.19, H: 5.02; N: 15.91.

Following the above described procedures, other cyclic esters are alsoprepared, such asN⁹-neopentyl-8-(2-(5-(1-(4-pyridyl)propan-1,3-yl)phosphono)furanyl)adenine,N⁹-neopentyl-8-(2-(5-(1-(3-pyridyl)propan-1,3-yl)phosphono)furanyl)adenine,andN⁹-neopentyl-8-(2-(5-(1-phenylpropan-1,3-yl)phosphono)furanyl)adenine.

Example 25 Preparation of 2-substituted-1,3-propanediol cyclic esters ofpurine phosphonates

Step A. To a solution of 2-(hydroxymethyl)-1,3-propanediol (1 g, 9.4mmol) in pyridine (7.5 mL) at 0° C. was added acetic anhydride (0.89 mL,9.4 mmol) slowly. The resulting solution was warmed to room temperatureand stirred for 16 h. The reaction was concentrated under reducedpressure and chromatographed by eluting with methanol-dichloromethane(1:9) to give 510 mg of pure 2-acetoxymethyl-1,3-propanediol.

Step B. 2-Acetoxymethyl-1,3-propanediol was coupled toN⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine following Step C ofExample 24 to giveN⁹-neopentyl-8-(2-(5-(2-(acetoxymethyl)propan-1,3-yl)phosphono)furanyl)adenine(25.1). mp=164-165° C.; Anal. Calcd. for C₂₀H₂₆N₅O₆P: C: 51.84; H: 5.65;N: 15.11. Found: C: 52.12; H: 5.77; N: 14.59.

Following the above described procedures, other cyclic esters are alsoprepared, such asN⁹-neopentyl-8-(2-(5-(2-(methoxycarbonyloxymethyl)-propan-1,3-yl)phosphono)furanyl)adenine,N⁹-neopentyl-8-(2-(5-(2-(hydroxymethyl)-propan-1,3-yl)phosphono)furanyl)adenine,N⁹-neopentyl-8-(2-(5-(2,2-dihydroxymethylpropan-1,3-yl)phosphono)furanyl)adenine,N⁹-neopentyl-8-(2-(5-(2-(methoxycarbonyloxymethyl)propan-1,3-yl)phosphono)-furanyl)adenineis prepared by coupling N⁹-neopentyl-8-(2-(5-phosphono)-furanyl)adeninewith 2-(methoxycarbonyloxymethyl)-1,3-propanediol which was prepared asfollows:

To a solution of 2-(hydroxymethyl)-1,3-propanediol (9.4 mmol) indichloromethane (20 mL) and pyridine (7.5 mL) at 0° C. was added methylchloroformate (9.4 mmol) slowly. The resulting solution was warmed toroom temperature and stirred for 16 h. The reaction was concentratedunder reduced pressure and chromatographed by eluting withmethanol-dichloromethane (1:4) to give 650 mg of2-(methoxycarbonyloxymethyl)-1,3-propanediol.

Example 26 Preparation of 8-(2-(5-hydroxyl-1,3cyclohexyl)phosphono)furanylpurines

A suspension of N⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine (1 mmol)in 5 mL of thionyl chloride was heated at reflux temperature for 4 h.The reaction mixture was cooled and evaporated to dryness. To theresulting residue was added a solution of cis,cis-1,3,5-cyclohexanetriol(1 mmol) and pyridine (2.5 mmol) in 3 mL of methylene chloride. Afterstirring at 25° C. for 24 h the reaction was subjected to work up andchromatography to give N⁹-neopentyl-8-(2-(5-(5-hydroxyl-1,3cyclohexyl)phosphono)furanyl)adenine, minor isomer (26.1). mp 248-250°C.; Anal. Cald. for C₂₀H₂₆N₅O₅P+0.5 H₂O: C: 52.63; H: 5.96; N: 15.34.Found: C: 52.62; H: 5.70; N: 15.32; major isomer (26.2). mp 225-230° C.;Anal. Cald. for C₂₀H₂₆N₅O₅P+0.5 H₂O: C: 52.63; H: 5.96; N: 15.34. Found:C: 52.74; H: 5.80; N: 15.32.

Following the above described procedures,N⁹-phenethyl-8-(2-(5-(5-hydroxyl-1,3cyclohexyl)phosphono)furanyl)adenine (26.3) was also prepared. Anal.Cald. for C₂₃H₂₄N₅O₅P+0.15 H₂O: C: 57.06; H: 5.06; N: 14.47. Found: C:56.84; H: 4.83; N: 14.38.

Example 27 Preparation of 3-(2-hydroxyethyl)phthalide

A solution of phthalide-3-acetic acid (1 mmol) in THF was treated withborane dimethylsulfide (1.5 mmol) at 0° C. for 1h, and 25° C. for 24 h.Extraction and chromatography gave 2-(3-phthalidyl)ethanol as a lightyellow oil. TLC: R_(f)=0.25, 50% EtOAc-hexane.

Example 28 Preparation of Purine Phosphonate Amine Salts

A mixture of N⁹-neopentyl-8-(2-(5-phosphono)furanyl)adenine (1 mmol) andtris(hydroxymethyl)aminomethane (1.05 mmol) in methanol is stirred at25° C. for 24 h. Evaporation givesN⁹-neopentyl-8-(2-(5-phosphono)furanyl)adeninetris(hydroxymethyl)aminomethane salt.

Examples of the methods of the present invention include the following.It will be understood that these examples are exemplary and that themethod of the invention is not limited solely to these examples.

For the purposes of clarity and brevity, chemical compounds in thefollowing biological examples are referred to by synthetic examplenumbers.

Besides the following Examples, assays that may be useful foridentifying compounds which inhibit gluconeogenesis include thefollowing animal models of Diabetes:

i. Animals with pancreatic b-cells destroyed by specific chemicalcytotoxins such as Alloxan or Streptozotocin (e.g. theStreptozotocin-treated mouse, -rat, dog, and -monkey). Kodama, H.,Fujita, M., Yamaguchi, I., Japanese Journal of Pharmacology 1994, 66,331-336 (mouse); Youn, J. H., Kim, J. K., Buchanan, T. A., Diabetes1994, 43, 564-571 (rat); Le Marchand, Y., Loten, E. G.,Assimacopoulos-Jannet, F., et al., Diabetes 1978, 27, 1182-88 (dog); andPitkin, R. M., Reynolds, W. A., Diabetes 1970, 19, 70-85 (monkey).

ii. Mutant mice such as the C57BUKs db/db, C57BUKs ob/ob, and C57BU6Job/ob strains from Jackson Laboratory, Bar Harbor, and others such asYellow Obese, T-KK, and New Zealand Obese. Coleman, D. L., Hummel, K.P., Diabetologia 1967, 3, 238-248 (C57BUKs db/db); Coleman, D. L.,Diabetologia 1978, 14,141-148 (C57BU6J ob/ob); Wolff, G. L., Pitot, H.C., Genetics 1973, 73,109-123 (Yellow Obese); Dulin, W. E., Wyse, B. M.,Diabetologia 1970, 6, 317-323 (T-KK); and Bielschowsky, M.,Bielschowsky, F. Proceedings of the University of Otago Medical School1953, 31, 29-31 (New Zealand Obese).

iii. Mutant rats such as the Zucker fa/fa Rat rendered diabetic withStreptozotocin or Dexamethasone, the Zucker Diabetic Fatty Rat, and theWistar Kyoto Fatty Rat. Stolz, K. J., Martin, R. J. Journal of Nutrition1982, 112, 997-1002 (Streptozotocin); Ogawa, A., Johnson, J. H.,Ohnbeda, M., McAllister, C. T., Inman, L., Alam, T., Unger, R. H., TheJournal of Clinical Investigation 1992, 90, 497-504 (Dexamethasone);Clark, J. B., Palmer, C. J., Shaw, W. N., Proceedings of the Society forExperimental Biology and Medicine 1983, 173, 68-75 (Zucker DiabeticFatty Rat); and Idida, H., Shino, A., Matsuo, T., et al., Diabetes 1981,30,1045-1050 (Wistar Kyoto Fatty Rat).

iv. Animals with spontaneous diabetes such as the Chinese Hamster, theGuinea Pig, the New Zealand White Rabbit, and non-human primates such asthe Rhesus monkey and Squirrel monkey. Gerritsen, G. C., Connel, M. A.,Blanks, M. C., Proceedings of the Nutrition Society 1981, 40, 237 245(Chinese Hamster); Lang, C. M., Munger, B. L., Diabetes 1976, 25,434-443 (Guinea Pig); Conaway, H. H., Brown, C. J., Sanders, L. L. etaI., Journal of Heredity 1980, 71, 179-186 (New Zealand White Rabbit);Hansen, B. C., Bodkin, M. L., Diabetologia 1986, 29, 713-719 (Rhesusmonkey); and Davidson, I. W., Lang, C. M., Blackwell, W. L., Diabetes1967, 16, 395-401 (Squirrel monkey).

v. Animals with nutritionally induced diabetes such as the Sand Rat, theSpiny Mouse, the Mongolian Gerbil, and the Cohen Sucrose-InducedDiabetic Rat. Schmidt-Nielsen, K., Hainess, H. B., Hackel, D. B.,Science 1964, 143, 689-690 (Sand Rat); Gonet, A. E., Stauffacher, W.,Pictet, R., et al., Diabetologia 1965, 1, 162-171 (Spiny Mouse);Boquist, L., Diabetologia 1972, 8, 274-282 (Mongolian Gerbil); andCohen, A. M., Teitebaum, A., Saliternik, R., Metabolism 1972, 21,235-240 (Cohen Sucrose-Induced Diabetic Rat).

vi. Any other animal with one of the following or a combination of thefollowing characteristics resulting from a genetic predisposition,genetic engineering, selective breeding, or chemical or nutritionalinduction: impaired glucose tolerance, insulin resistance,hyperglycemia, obesity, accelerated gluconeogenesis, increased hepaticglucose output.

Example A Inhibition of Human Liver FBPase

E. coli strain BL21 transformed with a human liver FBPase-encodingplasmid was obtained from Dr. M. R. El-Maghrabi at the State Universityof New York at Stony Brook. hIFBPase was typically purified from 10liters of E. coli culture as described (M. Gidh-Jain et al., The Journalof Biological Chemistry 1994, 269, 27732-27738). Enzymatic activity wasmeasured spectrophotometrically in reactions that coupled the formationof product (fructose 6-phosphate) to the reduction ofdimethylthiazoldiphenyltetrazolium bromide (MTT) via NADP and phenazinemethosulfate (PMS), using phosphoglucose isomerase and glucose6-phosphate dehydrogenase as the coupling enzymes. Reaction mixtures(200 μL) were made up in 96-well microtitre plates, and consisted of 50mM Tris-HCl, pH 7.4,100 mM KCl, 5 mM EGTA, 2 mM MgCl₂, 0.2 mM NADP, 1mg/mL BSA, 1 mM MTT, 0.6 mM PMS, 1 unit/mL phosphoglucose isomerase, 2units/mL glucose 6-phosphate dehydrogenase, and 0.150 mM substrate(fructose 1,6-bisphosphate). Inhibitor concentrations were varied from0.01 μM to 10 μM. Reactions were started by the addition of 0.002 unitsof pure hIFBPase and were monitored for 7 minutes at 590 nm in aMolecular Devices Plate Reader (37° C.).

The following Table depicts the IC₅₀ values for several compoundsprepared in the Examples. AMP has an IC₅₀ value of 1.0 μM in this assay.

Example Compound IC₅₀ (human Number liver FBPase 2.1 5 μM 2.2 1.4 μM 2.33.3 μM 2.7 0.8 μM 2.10 2 μM 2.13 4.5 μM 2.14 0.9 μM 2.16 1.4 μM 4.1 100μM 5.5 10 μM 8.1 23 μM 10.1 50 μM 11.1 0.7 μM 12.1 13 μM

FIG. 2 shows the dose-dependent inhibition of hIFBPase by AMP, compound2.7, compound 2.13, and compound 2.5.

In vitro Inhibition of Rat Liver and Mouse Liver FBPase Was AlsoDetermined

E. coil strain BL21 transformed with a rat liver FBPase-encoding plasmidwas obtained from Dr. M. R. EI-Maghrabi at the State University of NewYork at Stony Brook, and purified as described (EI-Maghrabi, M. R., andPilkis, S. J. Biochem. Biophys. Res. Commun. 1991, 176,137-144). Mouseliver FBPase was obtained by homogenizing freshly isolated mouse liverin 100 mM Tris-HCl buffer, pH 7.4, containing 1 mM EGTA, and 1 0%glycerol. The homogenate was clarified by centrifugation, and the 45-75%ammonium sulfate fraction prepared. This fraction was redissolved in thehomogenization buffer and desalted on a PD-10 gel filtration column(Biorad) eluted with same. This partially purified fraction was used forenzyme assays. Both rat liver and mouse liver FBPase were assayed asdescribed for human liver FBPase. Generally, as reflected by the higherIC₅₀ values, the rat and mouse liver enzymes are less sensitive toinhibition by the compounds tested than the human liver enzyme.

The following Table depicts the IC₅₀ values for several compoundsprepared in the Examples:

IC50 Rat Liver IC50 Mouse Liver Compound μM μM 2.1 100 >20 2.2 >20 >202.3 >20 >20 2.7 1.25 55 2.10 >20 >20 2.13 >20 >20 2.14 >20 >202.16 >20 >20 4.1 >20 >20 5.5 >20 >20 8.1 >20 >20 10.1 >20 >2011.1 >20 >20 12.1 20 >100

Example B AMP Site Binding

To determine whether compounds bind to the allosteric AMP binding siteof hIFBPase, the enzyme was incubated with radiolabeled AMP in thepresence of a range of test compound concentrations. The reactionmixtures consisted of 25 mM ³H-AMP (54 mCi/mmol) and 0-1000 mM testcompound in 25 mM Tris-HCl, pH 7.4,100 mM KCl and 1 mM MgCl₂. 1.45 mg ofhomogeneous FBPase (±1 nmole) was added last. After a 1 minuteincubation, AMP bound to FBPase was separated from unbound AMP by meansof a centrifugal ultrafiltration unit (“Ultrafree-MC”, Millipore) usedaccording to the instructions of the manufacturer. The radioactivity inaliquots (100 μl) of the upper compartment of the unit (the retentate,which contains enzyme and label) and the lower compartment (thefiltrate, which contains unbound label) were quantified using a Beckmanliquid scintillation counter. The amount of AMP bound to the enzyme wasestimated by comparing the counts in the filtrate (the unbound label) tothe total counts in the retentate.

As evident from FIG. 3, both aminoimidazolecarboxamideribosidemonophosphate (ZMP) and compound 2.2 displaced AMP from hIFBPase in adose-dependent manner, indicating that they bind to the same site on theenzyme as AMP. As expected, compound 2.2, a more potent hIHBPaseinhibitor than ZMP (IC₅₀'s=5 and 12 μM, respectively), had a lower ED₅₀for AMP displacement than ZMP (35 vs 250 μM).

Example C AMP Site/Enzyme Selectivity

To determine the selectivity of compounds towards FBPase, effects ofFBPase inhibitors on 5 key AMP binding enzymes were measured using theassays described below:

Adenosine Kinase:

Human adenosine kinase was purified from an E. coil expression system asdescribed by Spychala et al. (Spychala, J., Datta, N. S., Takabayashi,K., Datta, M., Fox, I. H., Gribbin, T., and Mitchell, B. S. Proc. Natl.Acad. Sci. USA 1996, 93, 1232-1237). Activity was measured essentiallyas described by Yamada et al. (Yamada, Y., Goto, H., Ogasawara, N.Biochim. Biophys. Acta 1988, 660, 36-43.) with a few minormodifications. Assay mixtures contained 50 mM TRIS-maleate buffer, pH7.0, 0.1% BSA, 1 mM ATP 1 mM MgCl₂, 1.0 μM [U-¹⁴C] adenosine (400-600mCi/mmol) and varying duplicate concentrations of inhibitor. ¹⁴C-AMP wasseparated from unreacted ¹⁴C-adenosine by absorption to anion exchangepaper (Whatman) and quantified by scintillation counting.

Adenosine Monophosphate Deaminase:

Porcine heart AMPDA was purified essentially as described by Smiley etal. (Smiley, K. L., Jr, Berry, A. J., and Suelter, C. H. J. Biol. Chem.1967, 242, 2502-2506) through the phosphocellulose step. Inhibition ofAMPDA activity was determined at 37° C. in a 0.1 mL assay mixturecontaining inhibitor, ˜0.005U AMPDA, 0.1% bovine serum albumin, 10 mMATP, 250 mM KCl, and 50 mM MOPS at pH 6.5. The concentration of thesubstrate AMP was varied from 0.125-10.0 mM. Catalysis was initiated bythe addition of enzyme to the otherwise complete reaction mixture, andterminated after 5 minutes by injection into an HPLC system. Activitieswere determined from the amount of IMP formed during 5 minutes. IMP wasseparated from AMP by HPLC using a Beckman Ultrasil-SAX anion exchangecolumn (4.6 mm×25 cm) with an isocratic buffer system (12.5 mM potassiumphosphate, 30 mM KCl, pH 3.5) and detected spectrophotometrically byabsorbance at 254 nm.

Phosphofructokinase:

Enzyme (rabbit liver) was purchased from Sigma. Activity was measured at30° C. in reactions in which the formation of fructose 1,6-bisphosphatewas coupled to the oxidation of NADH via the action of aldolase,triosephosphate isomerase, and α-glycerophosphate dehydrogenase.Reaction mixtures (200 μl) were made up in 96-well microtitre plates andwere read at 340 nm in a Molecular Devices Microplate Reader. Themixtures consisted of 200 mM Tris-HCl pH 7.0, 2 mM DTT, 2 mM MgCl₂, 0.2mM NADH, 0.2 mM ATP, 0.5 mM Fructose 6-phosphate, 1 unit aldolase/mL, 3units/mL triosephosphate isomerase, and 4 units/mL α-glycerophosphatedehydrogenase. Test compound concentrations ranged from 1 to 500 μM.Reactions were started by the addition of 0.0025 units ofphosphofructokinase and were monitored for 15 minutes.

Glycogen Phosphorylase:

Enzyme (rabbit muscle) was purchased from Sigma. Activity was measuredat 37° C. in reactions in which the formation of glucose 1-phosphate wascoupled to the reduction of NADP via phosphoglucomutase and glucose6-phosphate dehydrogenase. Assays were performed on 96-well microtitreplates and were read at 340 nm on a Molecular Devices Microplate Reader.Reaction mixtures consisted of 20 mM imidazole, pH 7.4, 20 mM MgCl₂, 150mM potassium acetate, 5 mM potassium phosphate, 1 mM DTT, 1 mg/mL BSA,0.1 mM NADP, 1 unit/mL phosphoglucomutase, 1 unit/mL glucose 6-phosphatedehydrogenase, 0.5% glycogen. Test compound concentrations ranged from 1to 500 μM. Reactions were started by the addition of 17 μg enzyme andwere monitored for 20 minutes.

Adenylate Kinase:

Enzyme (rabbit muscle) was purchased from Sigma. Activity was measuredat 37° C. in reaction mixtures (100 μl) containing 100 mM Hepes, pH 7.4,45 mM MgCl₂, 1 mM EGTA, 100 mM KCl, 2 mg/mL BSA, 1 mM AMP and 2 mM ATP.Reactions were started by addition of 4.4 ng enzyme and terminated after5 minutes by addition of 17 μl perchloric acid. Precipitated protein wasremoved by centrifugation and the supernatant neutralized by addition of33 μl 3 M KOH/3 M KH₂CO₃. The neutralized solution was clarified bycentrifugation and filtration and analyzed for ADP content (enzymeactivity) by HPLC using a YMC ODS AQ column (25×4.6 cm). A gradient wasrun from 0.1 M KH₂PO₄, pH 6, 8 mM tetrabutyl ammonium hydrogen sulfateto 75% acetonitrile. Absorbance was monitored at 254 nM.

Compound 2.1, a 5 μM hIFBPase inhibitor, was essentially inactive in allof the above described assays except for the AMP deaminase screen:half-maximal inhibition of AMP deaminase was observed at almost the sameconcentration as the IC₅₀ for FBPase. Compound 2.7 (hIFBPase IC₅₀=0.8μM), in addition to being essentially without effect on adenosinekinase, adenylate kinase, glycogen phosphorylase, andphosphofructokinase, was only a weak inhibitor of AMP deaminase(IC₅₀=390 μM). The data suggest that compound 2.7 binds to hIFBPase in ahighly selective manner. The following Table gives the selectivity datafor compounds 2.1 and 2.7.

SELECTIVITY Compound 2.1 Compound 2.7 FBPase (inh.) 5.0 μM 0.8 μMAdenosine Kinase (inh.) >>10 >>100 Adenylate Kinase (inh.) >>500 >>500AMP Deaminase (inh.) 6.7 390 Glycogen Phosphorylase (act.) >>250 >>100Phosphofructokinase (act.) >>200 >>100

Example D Inhibition of Gluconeogenesis in Rat Hepatocytes

Hepatocytes were prepared from overnight fasted Sprague-Dawley rats(250-300 g) according to the procedure of Berry and Friend (Berry, M.N., Friend, D. S., J. Cell. Biol. 1969, 43, 506-520) as modified byGroen (Groen, A. K., Sips, H. J., Vervoorn, R. C., Tager, J. M., Eur. J.Biochem. 1982, 122, 87-93). Hepatocytes (75 mg wet weight/mL) wereincubated in 1 mL Krebs-bicarbonate buffer containing 10 mM Lactate, 1mM pyruvate, 1 mg/mL BSA, and test compound concentrations from 1 to 500μM. Incubations were carried out in a 95% oxygen, 5% carbon dioxideatmosphere in closed, 50-mL Falcon tubes submerged in a rapidly shakingwater bath (37° C.). After 1 hour, an aliquot (0.25 mL) was removed,transferred to an Eppendorf tube and centrifuged. 50 μl of supernatantwas then assayed for glucose content using a Sigma Glucose Oxidase kitas per the manufacturer's instructions.

Compound 2.1 and compound 2.7 inhibited glucose production fromlactate/pyruvate in isolated rat hepatocytes in a dose-dependent manner,with IC₅₀'s of 90 and 4.5 μM, respectively (FIG. 5). IC₅₀'s for otherselect compounds in this assay are shown in the Table below:

Compound IC50 Glucose Production, μM 2.2 90 2.6 18 2.10 24 2.13 50 2.147.5 2.16 12 16.4 3

FPBase from rat liver is less sensitive to AMP than that from humanliver. IC₅₀ values are correspondingly higher in rat hepatocytes thanwould be expected in human hepatocytes.

Example E Effect of Compound 2.7 on Gluconeogenesis FromDihydroxyacetone in Rat Hepatocytes: Glucose Production Inhibition andFructose 1,6-bisphosphate Accumulation

Isolated rat hepatocytes were prepared as described in Example D andincubated under the identical conditions described except thatlactate/pyruvate was replaced by 10 mM dihydroxyacetone, a substratewhich feeds into the gluconeogenic pathway at a step just prior toFBPase. Reactions were terminated by removing an aliquot (250 μL) ofcell suspension and spinning it through a layer of oil (0.8 mLsilicone/mineral oil, 4/1) into a 10% perchloric acid layer (100 μL).After removal of the oil layer, the acidic cell extract layer wasneutralized by addition of ⅓rd volume of 3 M KOH/3 M KH₂CO₃. Afterthorough mixing and centrifugation, the supernatant was analyzed forglucose content as described in Example D, and also forfructose-1,6-bisphosphate. Fructose-1,6-bisphosphate was assayedspectrophotometrically by coupling its enzymatic conversion to glycerol3-phosphate to the oxidation of NADH, which was monitored at 340 nm.Reaction mixtures (1 mL) consisted of 200 mM Tris-HCl, pH 7.4, 0.3 mMNADH, 2 units/mL glycerol 3-phsophate dehydrogenase, 2 units/mLtriosephosphate isomerase, and 50-100 μl cell extract. After a 30 minutepreincubation at 37° C., 1 unit/mL of aldolase was added and the changein absorbance measured until a stable value was obtained. 2 moles ofNADH are oxidized in this reaction per mole of fructose-1,6-bisphosphatepresent in the cell extract.

As shown in FIG. 4A, compound 2.7 inhibited glucose production fromdihydroxyacetone in rat hepatocytes (IC₅₀ approx. 5 μM) as effectivelyas from lactate pyruvate (IC₅₀ 4.5 μM, FIG. 5). This data confirms thatthe site of action of the compound is in the last four steps of thegluconeogenic pathway. The dose-dependent accumulation offructose-1,6-bisphosphate (the substrate of FBPase) that occurs uponcell exposure to compound 2.7 (FIG. 4B) is consistent with theinhibition of FBPase, the second to last enzyme in the pathway.

Example F Blood Glucose Lowering in Fasted Rats

Sprague Dawley rats (250-300 g) were fasted for 18 hours and then dosedintraperitoneally either with saline or with 35, 45, and 60 mg/kgcompound 16.4, a prodrug of compound 2.7. The vehicle used for drugadministration was dimethylsulfoxide. Blood samples were obtained fromthe tail vein of conscious animals just prior to injection and then athalf-hourly intervals. Blood glucose was measured using a HemoCue Inc.glucose analyzer according to the instructions of the manufacturer.

FIG. 6 shows the profound glucose lowering elicited by treatment withcompound 16.4. The duration of action was dose-dependent and ranged from2 to 6 hours.

Example G Analysis of Drug Levels and Liver Fructose-1,6-bisphosphateAccumulation in Rats

Sprague-Dawley rats (250-300 g) were fasted for 18 hours and then dosedintraperitoneally either with saline (n=3) or 20 mgs/kg compound 2.7(n=4). The vehicle used for drug administration was 10 mM bicarbonate.One hour post injection rats were anesthetized with halothane and aliver biopsy (approx. 1 g) was taken as well as a blood sample (2 mL)from the posterior vena cava. A heparin flushed syringe and needle wereused for blood collection. The liver sample was immediately homogenizedin ice-cold 10% perchloric acid (3 mL), centrifuged, and the supernatantneutralized with ⅓rd volume of 3 M KOH/3 M KH₂CO₃. Followingcentrifugation and filtration, 50 μL of the neutralized extract wasanalyzed for compound 2.7 content by HPLC. A reverse phase YMC ODS AQcolumn (250×4.6 cm) was used and eluted with a gradient from 10 mMsodium phosphate pH 5.5 to 75% acetonitrile. Absorbance was monitored at310 nm. The concentration of fructose-1,6-bisphosphate in liver was alsoquantified using the method described in Example E. Blood glucose wasmeasured in the blood sample as described in Example F. Plasma was thenquickly prepared by centrifugation and extracted by addition of methanolto 60% (v/v). The methanolic extract was clarified by centrifugation andfiltration and then analyzed by HPLC as described above.

Compound 2.7 lowered blood glucose from 82±3 to 28±9.9 mg/dL within onehour (FIG. 7). Drug levels measured in plasma and liver were 38.5±7 μMand 51.3±10 nmoles/g, respectively. As shown in FIG. 8, a 10-foldelevation of fructose-1,6-bisphosphate levels was found in the liversfrom the drug-treated group, consistent with the inhibition of glucoseproduction at the level of FBPase in the gluconeogenic pathway.

Example H Blood Glucose Lowering in Zucker Diabetic Fatty Rats

Zucker Diabetic Fatty rats purchased at 7 weeks of age were used at age16 weeks in the 24-hour fasted state. The rats were purchased fromGenetics Models Inc. and fed the recommended Purina 5008 diet (6.5%fat). Their fasting hyperglycemia at 24 hours ranged from 150 mg/dL to310 mg/dL blood glucose.

Compound 2.7 was administered at a dose of 50 mg/kg by intraperitonealinjection (n=6). The stock solution was made up at 25 mg/mL in deionizedwater and adjusted to neutratility by dropwise addition of 5 N NaOH. 5control animals were dosed with saline. Blood glucose was measured atthe time of dosing and 2 hours post dose as described in Example F.

As shown in FIGS. 9A and 9B, blood glucose was lowered in thedrug-treated group by an average of almost 20% (p<0.0001 relative to thecontrol animals).

Example I Inhibition of Gluconeogenesis in Zucker Diabetic Fatty Rats

Three 20-week old Zucker Diabetic Fatty rats were dosed with compound2.7 and three with saline as described in Example H. Fifteen minutespost-injection, the animals were anesthetized with sodium pentobarbitol(30 mgs, i.p.) and ¹⁴C-bicarbonate (20 μCi/100 g of body weight) wasadministered via the tail vein. Blood samples (0.6 mL) were obtained bycardiac puncture 10 and 20 minutes post tracer injection. Blood (0.5 mL)was diluted into 6 mL deionized water and protein precipitated byaddition of 1 mL zinc sulfate (0.3 N) and 1 mL barium hydroxide (0.3 N). The mixture was centrifuged (20 minutes, 1000×g) and 5 mL of theresulting supernatant was then combined with 1 g of a mixed bed ionexchange resin (1 part AG 50W-X8, 100-200 mesh, hydrogen form and 2parts of AG 1-X8, 100-200 mesh, acetate form) to separate¹⁴C-bicarbonate from ¹⁴C-glucose. The slurry was shaken at roomtemperature for four hours and then allowed to settle. An aliquot of thesupernatant (0.5 mL) was then counted in 5 mL scintillation cocktail.

As shown in FIG. 10, compound 2.7 reduced the incorporation of¹⁴C-bicarbonate into glucose by 75%; therefore gluconeogenesis wasclearly inhibited by the drug.

Example J Blood Glucose Lowering in Streptozotocin-treated Rats

Diabetes is induced in male Sprague-Dawley rats (250-300g) byintraperitoneal injection of 55 mg/kg streptozotocin (Sigma ChemicalCo.). Six days later, 24 animals are selected with fed blood glucosevalues (8 am) between 350 and 600 mg/dL and divided into twostatistically equivalent groups. Blood glucose is measured in bloodobtained from a tail vein nick by means of a HemoCue Inc. (MissionViejo, Calif.) glucose analyzer. One group of 12 will subsequentlyreceive inhibitor (100 mg/kg intraperitoneally) and the other 12(“controls”) an equivalent volume of saline. Food is removed from theanimals. Blood glucose is measured in each animal four hours afterdosing, and a second dose of drug or saline is then administered. Fourhours later, a final blood glucose measurement is made.

Example K Evaluation of Compound 16.4 as a Prodrug in RatHepatocytes—Intracellular Delivery of Compound 2.7

Rat hepatocytes were prepared and incubated as in Example D, except thatthe test compound, 16.4, was added to yield a final concentration of 10μM. Aliquots of the cell suspension were taken at 0, 5, 10, 20, 30, 45,and 60 minutes after drug exposure. The cells were extracted andanalyzed for compound 2.7 content by HPLC as described in Example L.Absorbance of the HPLC column eluate was monitored at 310 nm.Quantitation of intracellular compound 2.7 was done by comparison toauthentic standards of known concentration. As shown in FIG. 11A,compound 16.4 rapidly delivered high levels of compound 2.7 into thehepatocytes; a concentration of approximately 80 nmoles/g was achievedwithin 10 minutes. These data indicate that compound 16.4 readilypenetrates cells and is efficiently de-esterified to the parentcompound, 2.7, intracellularly. Furthermore, as shown in FIG. 11B,compound 16.4 inhibited glucose production in rat hepatocytes.

Example L Estimation of the Oral Bioavailability of Prodrugs ofPhosphonic Acids

Prodrugs were dissolved in 10% ethanol/90% polyethylene glycol (mw 400)and administered by oral gavage at doses of approximately 20 or 40 mg/kgparent compound equivalents to 6-hour fasted, Sprague Dawley rats(220-240 g). The rats were subsequently placed in metabolic cages andurine was collected for 24 hours. The quantity of parent compoundexcreted into urine was determined by HPLC analysis. An ODS columneluted with a gradient from potassium phosphate buffer, pH 5.5 toacetonitrile was employed for these measurements. Detection was at310-325 nm. The percentage oral bioavailability was estimated bycomparison of the recovery in urine of the parent compound generatedfrom the prodrug, to that recovered in urine 24 hours after intravenousadministration of unsubstituted parent compound at approximately 10mg/kg. Parent compounds were typically dissolved in dimethyl sulfoxide,and administered via the tail vein in animals that were brieflyanesthetized with halothane.

For compound 16.4, a prodrug of compound 2.7, 6.2% of an oral dose ofapproximately 20 mg/kg was recovered in urine. For compound 2.7, 76.8%of an intravenous dose of approximately 10 mg/kg was recovered. The oralbioavailability of compound 16.4 was therefore calculated to be6.2/76.8, or approximately 8%. The oral bioavailability of compound 16.5was also estimated following the above described protocol to be 5.3%.

Example M Glucose Lowering Following Oral Administration of FBPaseInhibitors

FBPase inhibitor was administered by oral gavage at doses of 30, 100 and250 mg/kg to 18-hour fasted, Sprague Dawley rats (250-300g;n=4-5/group). The compound was prepared in deionized water, adjusted toneutrality with sodium hydroxide, and brought into solution bysonication prior to administration. Blood glucose was measuredimmediately prior to dosing, and at 1 hour intervals thereafter. Bloodsamples were obtained from the tail vein, and measurments made by meansof a Hemocue glucose analyzer (Hemocue Inc, Mission Viejo, Calif.) usedaccording to the manufacturer's instructions.

We claim:
 1. A compound of formula 1:

wherein A is selected from the group consisting of —NR⁸ ₂, —NHSO₂R³,—OR⁵, —SR⁵, halo, lower alkyl, —CON(R⁴)₂, guanidino, amidino, —H, andperhaloalkyl; E is selected from the group consisting of —H, halo, loweralkylthio, lower perhaloalkyl, lower alkyl, lower alkenyl, loweralkynyl, lower alkoxy, —CN, and —NR⁷ ₂; X is selected from the groupconsisting of -alk-NR—, alkylene, alkenylene, alkynylene, arylene,heteroarylene, -alk-NR-alk-, -alk-O-alk-, -alk-S-alk-, -alk-S—,alicyclicene, heteroalicyclicene, 1,1-dihaloalkylene, —C(O)-alk-,—NR—C(O)—NR′—, -alk-NR—C(O)—, -alk-C(O)—NR—, -Ar-alk-, and -alk-Ar-, alloptionally substituted, wherein each R and R′ is independently selectedfrom —H and lower alkyl, and wherein each “alk” and “Ar” is anindependently selected alkylene or arylene, respectively; Y is selectedfrom the group consisting of —H, alkyl, alkenyl, alkynyl, aryl,alicyclic, heteroalicyclic, aralkyl, aryloxyalkyl, alkoxyalkyl, —C(O)R³,—S(O)₂R³, —C(O)—OR³, —CONHR³, —NR² ₂, and —OR³, all except H areoptionally substituted; R¹ is independently selected from the groupconsisting of —H, alkyl, aryl, heteroalicyclic where the cyclic moietycontains a carbonate or thiocarbonate, —C(R²)₂-aryl, -alk-aryl,—C(R²)₂OC(O)NR² ₂, —NR²—C(O)—R³, —C(R²)₂—OC(O)R³, —C(R²)₂—O—C(O)OR³,—C(R²)₂OC(O)SR³, -alk-S—C(O)R³, -alk-S—S-alkylhydroxy, and-alk-S—S—S-alkylhydroxy, or together R¹ and R¹ are -alk-S—S-alk- to forma cyclic group, wherein each “alk” is an independently selectedalkylene, or together R¹ and R¹ are

wherein V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or together V and Z are connected via achain of 3-5 atoms, only one of which can be a heteroatom, to form partof a cyclic group substituted with hydroxy, acyloxy, alkoxycarboxy, oraryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; or together V and W are connected viaa chain of 3 carbon atoms to form part of a cyclic group substitutedwith hydroxy, acyloxy, alkoxycarboxy, alkylthiocarboxy, hydroxymethyl,or aryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; Z is selected from the groupconsisting of —CH₂OH, —CH₂OCOR³, —CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³,—S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar, —CH(Ar)OH, —CH(CH═CR²R²)OH,—CH(C≡CR²)OH, and —R²; with the provisos that: a) V, Z, W are not all—H; and b) when Z is —R², then at least one of V and W is not —H or —R⁹;R₂ is selected from the group consisting of R³ and —H; R³ is selectedfrom the group consisting of alkyl, aryl, alicyclic, heteroalicyclic,and aralkyl; R⁴ is independently selected from the group consisting of—H, lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,and lower aryl; R⁵ is selected from the group consisting of lower alkyl,lower aryl, lower aralkyl, lower alicyclic, and lower heteroalicyclic;R⁶ is independently selected from the group consisting of —H, and loweralkyl; R⁷ is independently selected from the group consisting of —H,lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,lower aryl, and —C(O)R¹⁰; R⁸ is independently selected from the groupconsisting of —H, lower alkyl, lower aralkyl, lower aryl, loweralicyclic, —C(O)R¹⁰, or together said R⁸ groups form a bidendatealkylene; R⁹ is selected from the group consisting of alkyl, aralkyl,alicyclic, and heteroalicyclic; R¹⁰ is selected from the groupconsisting of —H, lower alkyl, —NH₂, lower aryl, and lower perhaloalkyl;R¹¹ is selected from the group consisting of alkyl, aryl, —OH, —NH₂ and—OR³; and pharmaceutically acceptable prodrugs and salts thereof.
 2. Thecompounds of claim 1 with the proviso that R¹ is not lower alkyl of 1-4carbon atoms.
 3. The compounds of claim 1 wherein A is selected from thegroup consisting of —NR⁸ ₂, halo, lower alkyl, lower perhaloalkyl, andlower alkoxy.
 4. The compounds of claim 1 wherein E is —H, halo, lowerperhaloalkyl, —CN, lower alkyl, lower alkoxy, and lower alkylthio. 5.The compounds of claim 1 wherein X is selected from the group consistingof -alk-NR—, alkylene, alkynylene, -alk-O-alk-, -alk-S—, arylene,heteroarylene 1,1-dihaloalkylene, —C(O)-alk-, heteroarylene,-alk-C(O)—NR—, and -alk-NR—C(O)—, wherein each R is independentlyselected from —H and lower alkyl and wherein each “alk” is anindependently selected alkylene.
 6. The compounds of claim 1 wherein Xis alkylene substituted with 1 to 3 substituents selected from the groupconsisting of halo, —CO₂H, and —OH.
 7. The compounds of claim 5 whereinX is selected from the group consisting of -alk-NR—C(O)—, -alk-O-alk-,and heteroarylene.
 8. The compounds of claim 7 wherein X is selectedfrom the group consisting of methyleneoxymethylene and optionallysubstituted furandiyl.
 9. The compounds of claim 1 wherein Y is selectedfrom the group consisting of aralkyl, aryl, alicyclic, heteroalicyclic,and alkyl.
 10. The compounds of claim 1 wherein each R¹ is independentlyselected from the group consisting of —H, alkyl, aryl, heteroalicyclicwhere the cyclic moiety contains a carbonate or thiocarbonate,optionally substituted phenyl, optionally substituted benzyl, optionallysubstituted -alk-aryl, —C(R²)₂OC(O)R³, —C(R²)₂—O—C(O)OR³,—C(R²)₂—OC(O)SR³, -alk-S—C(O)R³, -alk-S—S-alkylhydroxy, and-alk-S—S—S-alkylhydroxy, or together R¹ and R¹ are -alk-S—S-alk- to forma cyclic group, wherein each “alk” is an independently selectedalkylene, or R¹ and R¹ together are

wherein V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or together V and Z are connected via achain of 3-5 atoms, only one of which can be a heteroatom, to form partof a cyclic group substituted with hydroxy, acyloxy, alkoxycarboxy, oraryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; or together V and W are connected viaa chain of 3 carbon atoms to form part of a cyclic group substitutedwith hydroxy, acyloxy, alkoxycarboxy, alkylthiocarboxy, hydroxymethyl,or aryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; Z is selected from the groupconsisting of —CH₂OH, —CH₂OCOR³, —CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³,—S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar, —CH(Ar)OH, —CH(CH═CR²R²)OH,—CH(C≡CR²)OH, and —R²; with the provisos that: a) V, Z, W are not all—H; and b) when Z is —R², then at least one of V and W is not —H or —R⁹;R₂ is selected from the group consisting of R³ and —H; R³ is selectedfrom the group consisting of alkyl, aryl, alicyclic, heteroalicyclic,and aralkyl; and R⁹ is selected from the group consisting of alkyl,aralkyl, alicyclic, and heteroalicyclic.
 11. The compounds of claim 10wherein each R¹ is independently selected from the group consisting ofoptionally substituted phenyl, optionally substituted benzyl,—C(R²)₂OC(O)R³, —C(R²)₂OC(O)OR³, and —H.
 12. The compounds of claim 10wherein R¹ is H.
 13. The compounds of claim 10 wherein at least one R¹is aryl, or —C(R²)₂-aryl.
 14. The compounds of claim 10 wherein at leastone R¹ is —C(R²)₂—OC(O)R³, —C(R²)₂—OC(O)OR³, —C(R²)₂—OC(O)SR³.
 15. Thecompounds of claim 10 wherein at least one R¹ is -alk-S—S-alkylhydroxy,-alk-S—C(O)R³, and -alk-S—S—S-alkylhydroxy, or together R¹ and R¹ are-alk-S—S-alk- to form a cyclic group, wherein each “alk” is anindependently selected alkylene.
 16. The compounds of claim 10 whereintogether R¹ and R¹ are

to form a cyclic group; wherein V and W are independently selected fromthe group consisting of hydrogen, aryl, substituted aryl, heteroaryl,substituted heteroaryl, 1-alkenyl, 1-alkynyl, and —R⁹; or together V andZ are connected via a chain of 3-5 atoms, only one of which can be aheteroatom, to form part of a cyclic group, substituted with hydroxy,acyloxy, alkoxycarboxy, or aryloxycarboxy attached to a carbon atom thatis three atoms from an oxygen attached to the phosphorus; or together Vand W are connected via a chain of 3 carbon atoms to form part of acyclic group substituted with hydroxy, acyloxy, alkoxycarboxy,alkylthiocarboxy, hydroxymethyl, or aryloxycarboxy attached to a carbonatom that is three atoms from an oxygen attached to the phosphorus; Z isselected from the group consisting of —CH₂OH, —CH₂OCOR³, —CH₂OC(O)SR³,—CH₂OCO₂R³, —SR³, —S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar, —CH(Ar)OH,—CH(CH═CR²R²)OH, —CH(C≡CR²)OH, and —R²; with the provisos that: a) V, Z,W are not all —H; and b) when Z is —R², then at least one of V and W isnot —H or —R⁹; R² is selected from the group consisting of R³ and —H; R³is selected from the group consisting of alkyl, aryl, alicyclic,heteroalicyclic, and aralkyl; and R⁹ is selected from the groupconsisting of alkyl, aralkyl, alicyclic, and heteroalicyclic.
 17. Thecompounds of claim 16 wherein V and W both form a 6-membered carbocyclicring substituted with 0-4 groups, selected from the group consisting ofhydroxy, acyloxy, alkoxycarbonyloxy, and alkoxy; and Z is R².
 18. Thecompounds of claim 16 wherein V and W are hydrogen; and Z is selectedfrom the group consisting of hydroxyalkyl, acyloxyalkyl, alkyloxyalkyl,and alkoxycarboxyalkyl.
 19. The compounds of claim 16 wherein V and Ware independently selected from the group consisting of hydrogen,optionally substituted aryl, and optionally substituted heteroaryl, withthe proviso that at least one of V and W is optionally substituted arylor optionally substituted heteroaryl.
 20. A method of treating an animalfor a disease derived from abnormally elevated insulin levels,comprising administering to said animal a therapeutically effectiveamount of a fructose-1,6-bisphosphatase inhibitor wherein said inhibitoris a compound of formula (1):

wherein A is selected from the group consisting of —NR⁸ ₂, NHSO₂R³,—OR⁵, —SR⁵, halo, lower alkyl, —CON(R⁴)₂, guanidino, amidino, —H, andperhaloalkyl; E is selected from the group consisting of —H, halo, loweralkylthio, lower perhaloalkyl, lower alkyl, lower alkenyl, loweralkynyl, lower alkoxy, —CN, and —NR⁷ ₂; X is selected from the groupconsisting of -alk-NR—, alkylene, alkenylene, alkynylene, arylene,heteroarylene, -alk-NR-alk-, -alk-O-alk-, -alk-S-alk-, -alk-S—,alicyclicene, heteroalicyclicene, 1,1-dihaloalkylene, —C(O)-alk-,—NR—C(O)—NR′—, -alk-NR—C(O)—, -alk-C(O)—NR—, -Ar-alk-, and -alk-Ar-, alloptionally substituted, wherein each R and R′ is independently selectedfrom —H and lower alkyl, and wherein each “alk” and “Ar” is anindependently selected alkylene or arylene, respectively; Y is selectedfrom the group consisting of —H, alkyl, alkenyl, alkynyl, aryl,alicyclic, heteroalicyclic, aralkyl, aryloxyalkyl, alkoxyalkyl, —C(O)R³,—S(O)₂R³, —C(O)—OR³, —CONHR³, —NR² ₂, and —OR³, all except H areoptionally substituted; R¹ is independently selected from the groupconsisting of —H, alkyl, aryl, heteroalicyclic where the cyclic moietycontains a carbonate or thiocarbonate, —C(R²)₂-aryl, -alk-aryl,—C(R²)₂OC(O)NR² ₂, —NR²—C(O)—R³, —C(R²)₂—OC(O)R³, —C(R²)₂—O—C(O)OR³,—C(R²)₂OC(O)SR³, -alk-S—C(O)R³, -alk-S—S-alkylhydroxy, and-alk-S—S—S-alkylhydroxy, or together R¹ and R¹ are -alk-S—S-alk- to forma cyclic group, wherein each “alk” is an independently selectedalkylene, or together R¹ and R¹ are

wherein V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or together V and Z are connected via achain of 3-5 atoms, only one of which can be a heteroatom, to form partof a cyclic group substituted with hydroxy, acyloxy, alkoxycarboxy, oraryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; or together V and W are connected viaa chain of 3 carbon atoms to form part of a cyclic group substitutedwith hydroxy, acyloxy, alkoxycarboxy, alkylthiocarboxy, hydroxymethyl,or aryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; Z is selected from the groupconsisting of —CH₂OH, —CH₂OCOR³, —CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³,—S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar, —CH(Ar)OH, —CH(CH═CR²R²)OH,—CH(C≡CR²)OH, and —R²; with the provisos that: a) V, Z, W are not all—H; and b) when Z is —R², then at least one of V and W is not —H or —R⁹;R₂ is selected from the group consisting of R³ and —H; R³ is selectedfrom the group consisting of alkyl, aryl, alicyclic, heteroalicyclic,and aralkyl; R⁴ is independently selected from the group consisting of—H, lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,and lower aryl; R⁵ is selected from the group consisting of lower alkyl,lower aryl, lower aralkyl, lower alicyclic, and lower heteroalicyclic;R⁶ is independently selected from the group consisting of —H, and loweralkyl; R⁷ is independently selected from the group consisting of —H,lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,lower aryl, and —C(O)R¹⁰; R⁸ is independently selected from the groupconsisting of —H, lower alkyl, lower aralkyl, lower aryl, loweralicyclic, —C(O)R¹⁰, or together said R⁸ groups form a bidendatealkylene; R⁹ is selected from the group consisting of alkyl, aralkyl,alicyclic, and heteroalicyclic; R¹⁰ is selected from the groupconsisting of —H, lower alkyl, —NH₂, lower aryl, and lower perhaloalkyl;R¹¹ is selected from the group consisting of alkyl, aryl, —OH, —NH₂ and—OR³; and pharmaceutically acceptable prodrugs and salts thereof. 21.The compounds of claim 10 wherein R¹ is heteroalicyclic where the cyclicmoiety contains carbonate or thiocarbonate.
 22. The compounds of claim21 wherein each R¹ is independently an optionally substituted2-oxo-1,3-dioxolene-4-yl attached through a methylene to the phosphorusoxygen.
 23. The compounds of claim 1 wherein A is selected from thegroup consisting of —NR⁸ ₂, and halo; E is selected from the groupconsisting of —H, halo, —CN, lower alkyl, lower perhaloalkyl, loweralkoxy, and lower alkylthio; X is selected from the group consisting of-alk-NR—, alkylene, -alk-O-alk-, alkynylene, -alk-S—, arylene,heteroarylene, -alk-C(O)—NR—, 1,1-dihaloalkylene, —C(O)-alk-, -alk(OH)—,and -alk-NR—C(O)—, wherein each R is independently selected from —H andlower alkyl, and wherein each “alk” is an independently selectedalkylene; and R⁴ and R⁷ are selected from the group consisting of —H,and lower alkyl.
 24. The compounds of claim 23 wherein Y is selectedfrom the group consisting of aralkyl, aryl, alicyclic, heteroalicyclic,and alkyl.
 25. The compound of claim 24 wherein R¹ and R¹ together are

V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or together V and Z are connected via achain of 3-5 atoms, only one of which can be a heteroatom, to form partof a cyclic group, substituted with hydroxy, acyloxy, alkoxycarboxy, oraryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; or together V and W are connected viaa chain of 3 carbon atoms to form part of a cyclic group, substitutedwith hydroxy, acyloxy, alkoxycarboxy, alkylthiocarboxy, hydroxymethyl,or aryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; Z is selected from the groupconsisting of —CH₂OH, —CH₂OCOR³, —CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³,—S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar, —CH(Ar)OH, —CH(CH═CR²R²)OH,—CH(C≡CR²)OH, and —R²; with the provisos that: a) V, Z, W are not all—H; and b) when Z is —R², then at least one of V and W is not —H or —R⁹;R² is selected from the group consisting of R³ and —H; R³ is selectedfrom the group consisting of alkyl, aryl, alicyclic, heteroalicyclic,and aralkyl; and R⁹ is selected from the group consisting of alkyl,aralkyl, alicyclic, and heteroalicyclic.
 26. The compounds of claim 23wherein A is —NR⁸ ₂; E is selected from the group consisting of —H, —Cl,and —SCH₃; and X is selected from the group consisting of optionallysubstituted furandiyl and -alk-O-alk-, wherein each “alk” is anindependently selected alkylene.
 27. The compounds of claim 26 wherein Ais —NH₂; E is selected from the group consisting of —H, —Cl, and —SCH₃;X is selected from the group consisting of 2,5-furandiyl andmethyleneoxymethylene; and Y is lower alkyl.
 28. The compound of claim27 wherein E is —H, X is 2,5-furandiyl, and Y is neopentyl.
 29. Thecompound of claim 28 wherein R¹ is —CH₂O—C(O)—C(CH₃)₃ or its HCl salt.30. The compound of claim 27 wherein E is —SCH₃, X is 2,5-furandiyl, andY is isobutyl.
 31. The compound of claim 30 wherein R¹ is—CH₂O—C(O)—C(CH₃)₃ or its HCl salt.
 32. The compound of claim 27 whereinE is —H, X is 2,5 furandiyl, and Y is 1-(3-chloro-2,2-dimethyl)-propyl.33. The compound of claim 32 wherein R¹ is —CH₂O—C(O)—C(CH₃)₃ or its HClsalt.
 34. A method of treating an animal for diabetes mellitus,comprising administering to said animal a therapeutically effectiveamount of a compound of formula (1):

wherein A is selected from the group consisting of —NR⁸ ₂, NHSO₂R³,—OR⁵, —SR⁵, halo, lower alkyl, —CON(R⁴)₂, guanidino, amidino, —H, andperhaloalkyl; E is selected from the group consisting of —H, halo, loweralkylthio, lower perhaloalkyl, lower alkyl, lower alkenyl, loweralkynyl, lower alkoxy, —CN, and —NR⁷ ₂; X is selected from the groupconsisting of -alk-NR—, alkylene, alkenylene, alkynylene, arylene,heteroarylene, -alk-NR-alk-, -alk-O-alk-, -alk-S-alk-, -alk-S—,alicyclicene, heteroalicyclicene, 1,1-dihaloalkylene, —C(O)-alk-,—NR—C(O)—NR′—, -alk-NR—C(O)—, -alk-C(O)—NR—, -Ar-alk-, and -alk-Ar-, alloptionally substituted, wherein each R and R′ is independently selectedfrom —H and lower alkyl, and wherein each “alk” and “Ar” is anindependently selected alkylene or arylene, respectively; Y is selectedfrom the group consisting of —H, alkyl, alkenyl, alkynyl, aryl,alicyclic, heteroalicyclic, aralkyl, aryloxyalkyl, alkoxyalkyl, —C(O)R³,—S(O)₂R³, —C(O)—OR³, —CONHR³, —NR² ₂, and —OR³, all except H areoptionally substituted; R¹ is independently selected from the groupconsisting of —H, alkyl, aryl, heteroalicyclic where the cyclic moietycontains a carbonate or thiocarbonate, —C(R²)₂-aryl, -alk-aryl,—C(R²)₂OC(O)NR² ₂, —NR²—C(O)—R³, —C(R²)₂—OC (O)R³, —C(R²)₂—O—C(O)OR³,—C(R²⁾ ₂OC(O)SR³, -alk-S—C(O)R³, -alk-S—S-alkylhydroxy, and-alk-S—S—S-alkylhydroxy, or together R¹ and R¹ are -alk-S—S-alk- to forma cyclic group, wherein each “alk” is an independently selectedalkylene, or together R¹ and R¹ are

wherein V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or together V and Z are connected via achain of 3-5 atoms, only one of which can be a heteroatom, to form partof a cyclic group substituted with hydroxy, acyloxy, alkoxycarboxy, oraryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; or together V and W are connected viaa chain of 3 carbon atoms to form part of a cyclic group substitutedwith hydroxy, acyloxy, alkoxycarboxy, alkylthiocarboxy, hydroxymethyl,or aryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; Z is selected from the groupconsisting of —CH₂OH, —CH₂OCOR³, —CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³,—S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar, —CH(Ar)OH, —CH(CH═CR²R²)OH,—CH(C≡CR²)OH, and —R²; with the provisos that: a) V, Z, W are not all—H; and b) when Z is —R², then at least one of V and W is not —H or —R⁹;R² is selected from the group consisting of R³ and —H; R³ is selectedfrom the group consisting of alkyl, aryl, alicyclic, heteroalicyclic,and aralkyl; R⁴ is independently selected from the group consisting of—H, lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,and lower aryl; R⁵ is selected from the group consisting of lower alkyl,lower aryl, lower aralkyl, lower alicyclic, and lower heteroalicyclic;R⁶ is independently selected from the group consisting of —H, and loweralkyl; R⁷ is independently selected from the group consisting of —H,lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,lower aryl, and —C(O)R¹⁰; R⁸ is independently selected from the groupconsisting of —H, lower alkyl, lower aralkyl, lower aryl, loweralicyclic, —C(O)R¹⁰, or together said R⁸ groups form a bidendatealkylene; R⁹ is selected from the group consisting of alkyl, aralkyl,alicyclic, and heteroalicyclic; R¹⁰ is selected from the groupconsisting of —H, lower alkyl, —NH₂, lower aryl, and lower perhaloalkyl;R¹¹ is selected from the group consisting of alkyl, aryl, —OH, —NH₂ and—OR³; and pharmaceutically acceptable prodrugs and salts thereof.
 35. Amethod of lowering blood glucose levels in an animal in need thereof,comprising administering to said animal a pharmaceutically acceptableamount of a compound of formula (1):

wherein A is selected from the group consisting of —NR⁸ ₂, NHSO₂R³,—OR⁵, —SR⁵, halo, lower alkyl, —CON(R⁴)₂, guanidino, amidino, —H, andperhaloalkyl; E is selected from the group consisting of —H, halo, loweralkylthio, lower perhaloalkyl, lower alkyl, lower alkenyl, loweralkynyl, lower alkoxy, —CN, and —NR⁷ ₂; X is selected from the groupconsisting of -alk-NR—, alkylene, alkenylene, alkynylene, arylene,heteroarylene, -alk-NR-alk-, -alk-O-alk-, -alk-S-alk-, -alk-S—,alicyclicene, heteroalicyclicene, 1,1-dihaloalkylene, —C(O)-alk-,—NR—C(O)—NR′—, -alk-NR—C(O)—, -alk-C(O)—NR—, -Ar-alk-, and -alk-Ar-, alloptionally substituted wherein each R and R¹ is independently selectedfrom —H and lower alkyl, and wherein each “alk” and “Ar” is anindependently selected alkylene or arylene, respectively; Y is selectedfrom the group consisting of —H, alkyl, alkenyl, alkynyl, aryl,alicyclic, heteroalicyclic, aralkyl, aryloxyalkyl, alkoxyalkyl, —C(O)R³,—S(O)₂R³, —C(O)—OR³, —CONHR³, —NR² ₂, and —OR³, all except H areoptionally substituted; R¹ is independently selected from the groupconsisting of —H, alkyl, aryl, heteroalicyclic where the cyclic moietycontains a carbonate or thiocarbonate, —C(R²)₂-aryl, -alk-aryl,—C(R²)₂OC(O)NR² ₂, —NR²—C(O)—R³, —C(R²)₂—OC (O)R³, —C(²)₂—O—C(O)OR³,—C(R²)₂OC(O)SR³, -alk-S—C(O)R³, -alk-S—S-alkylhydroxy, and-alk-S—S—S-alkylh droxy, or together R¹ and R¹ are -alk-S—S-alk- to forma cyclic group, wherein each “alk” is an independently selectedalkylene, or together R¹ and R¹ are

wherein V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or together V and Z are connected via achain of 3-5 atoms, only one of which can be a heteroatom, to form partof a cyclic group substituted with hydroxy, acyloxy, alkoxycarboxy, oraryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; or together V and W are connected viaa chain of 3 carbon atoms to form part of a cyclic group substitutedwith hydroxy, acyloxy, alkoxycarboxy, alkylthiocarboxy, hydroxymethyl,or aryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; Z is selected from the groupconsisting of —CH₂OH, —CH₂OCOR³, —CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³,—S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar, —CH(Ar)OH, —CH(CH═CR²R²)OH,—CH(C≡CR²)OH, and —R²; with the provisos that: a) V, Z, W are not all—H; and b) when Z is —R², then at least one of V and W is not —H or —R⁹;R² is selected from the group consisting of R³ and —H; R³ is selectedfrom the group consisting of alkyl, aryl, alicyclic, heteroalicyclic,and aralkyl; R⁴ is independently selected from the group consisting of—H, lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,and lower aryl; R⁵ is selected from the group consisting of lower alkyl,lower aryl, lower aralkyl, lower alicyclic, and lower heteroalicyclic;R⁶ is independently selected from the group consisting of —H, and loweralkyl; R⁷ is independently selected from the group consisting of —H,lower alkyl lower alicyclic, lower heteroalicyclic, lower aralkyl, loweraryl, and —C(O)R¹⁰; R⁸ is independently selected from the groupconsisting of —H, lower alkyl, lower aralkyl, lower aryl, loweralicyclic, —C(O)R¹⁰, or together said R⁸ groups form a bidendatealkylene; R⁹ is selected from the group consisting of alkyl, aralkyl,alicyclic, and heteroalicyclic; R¹⁰ is selected from the groupconsisting of —H, lower alkyl, —NH₂, lower aryl, and lower perhaloalkyl;R¹¹ is selected from the group consisting of alkyl, aryl, —OH, —NH₂ and—OR³; and pharmaceutically acceptable prodrugs and salts thereof.
 36. Amethod of treating an animal with excess glycogen storage disease,comprising administering to said animal in need thereof atherapeutically effective amount of a fructose-1,6-bisphosphataseinhibitor, wherein said inhibitor is a compound of formula (1):

wherein A is selected from the group consisting of —NR⁸ ₂, NHSO₂R³,—OR⁵, —SR⁵, halo, lower alkyl, —CON(R⁴)₂, guanidino, amidino, —H, andperhaloalkyl; E is selected from the group consisting of —H, halo, loweralkylthio, lower perhaloalkyl, lower alkyl, lower alkenyl, loweralkynyl, lower alkoxy, —CN, and —NR⁷ ₂; X is selected from the groupconsisting of -alk-NR—, alkylene, alkenylene, alkynylene, arylene,heteroarylene, -alk-NR-alk-, -alk-O-alk-, -alk-S-alk-, -alk-S—,alicyclicene, heteroalicyclicene, 1,1-dihaloalkylene, —C(O)-alk-,—NR—C(O)—NR′—, -alk-NR—C(O)—, -alk-C(O)—NR—, -Ar-alk-, and -alk-Ar-, alloptionally substituted, wherein each R and R′ is independently selectedfrom —H and lower alkyl, and wherein each “alk” and “Ar” is anindependently selected alkylene or arylene, respectively; Y is selectedfrom the group consisting of —H, alkyl, alkenyl, alkynyl, aryl,alicyclic, heteroalicyclic, aralkyl, aryloxyalkyl, alkoxyalkyl, —C(O)R³,—S(O)₂R³, —C(O)—OR³, —CONHR³, —NR² ₂, and —OR³, all except H areoptionally substituted; R¹ is independently selected from the groupconsisting of —H, alkyl, aryl, heteroalicyclic where the cyclic moietycontains a carbonate or thiocarbonate, —C(R²)₂-aryl, -alk-aryl,—C(R²)₂OC(O)NR² ₂, —NR²—C(O)—R³, —C(R²)₂—OC(O)R³, —C(R²)₂—O—C(O)OR³,—C(R²)₂OC(O)SR³, -alk-S—C(O)R³, -alk-S—S-alkylhydroxy, and-alk-S—S—S-alkylhydroxy, or together R¹ and R¹ are -alk-S—S-alk- to forma cyclic group, wherein each “alk” is an independently selectedalkylene, or together R¹ and R¹ are

wherein V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or together V and Z are connected via achain of 3-5 atoms, only one of which can be a heteroatom, to form partof a cyclic group substituted with hydroxy, acyloxy, alkoxycarboxy, oraryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; or together V and W are connected viaa chain of 3 carbon atoms to form part of a cyclic group substitutedwith hydroxy, acyloxy, alkoxycarboxy, alkylthiocarboxy, hydroxymethyl,or aryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; Z is selected from the groupconsisting of —CH₂OH, —CH₂OCOR³, —CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³,—S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar, —CH(Ar)OH, —CH(CH═CR²R²)OH,—CH(C≡CR²)OH, and —R²; with the provisos that: a) V, Z, W are not all—H; and b) when Z is —R², then at least one of V and W is not —H or —R⁹;R² is selected from the group consisting of R³ and —H; R³ is selectedfrom the group consisting of alkyl, aryl, alicyclic, heteroalicyclic,and aralkyl; R⁴ is independently selected from the group consisting of—H, lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,and lower aryl; R⁵ is selected from the group consisting of lower alkyl,lower aryl, lower aralkyl, lower alicyclic, and lower heteroalicyclic;R⁶ is independently selected from the group consisting of —H, and loweralkyl; R⁷ is independently selected from the group consisting of —H,lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,lower aryl, and —C(O)R¹⁰; R⁸ is independently selected from the groupconsisting of —H, lower alkyl, lower aralkyl, lower aryl, loweralicyclic, —C(O)R¹⁰, or together said R⁸ groups form a bidendatealkylene; R⁹ is selected from the group consisting of alkyl, aralkyl,alicyclic, and heteroalicyclic; R¹⁰ is selected from the groupconsisting of —H, lower alkyl, —NH₂, lower aryl, and lower perhaloalkyl;R¹¹ is selected from the group consisting of alkyl, aryl, —OH, —NH₂ and—OR³; and pharmaceutically acceptable prodrugs and salts thereof.
 37. Amethod of inhibiting gluconeogenesis in animal in need thereof,comprising administering to said animal an effective amount of acompound of formula (1):

wherein A is selected from the group consisting of —NR⁸ ₂, NHSO₂R³,—OR⁵, —SR⁵, halo, lower alkyl, —CON(R⁴)₂, guanidino, amidino, —H, andperhaloalkyl; E is selected from the group consisting of —H, halo, loweralkylthio, lower perhaloalkyl, lower alkyl, lower alkenyl, loweralkynyl, lower alkoxy, —CN, and —NR⁷ ₂; X is selected from the groupconsisting of -alk-NR—, alkylene, alkenylene, alkynylene, arylene,heteroarylene, -alk-NR-alk-, -alk-O-alk-, -alk-S-alk-, -alk-S—,alicyclicene, heteroalicyclicene, 1,1-dihaloalkylene, —C(O)-alk-,—NR—C(O)—NR′—, -alk-NR—C(O)—, -alk-C(O)—NR—, -Ar-alk-, and -alk-Ar-, alloptionally substituted, wherein each R and R′ is independently selectedfrom —H and lower alkyl, and wherein each “alk” and “Ar” is anindependently selected alkylene or arylene, respectively; Y is selectedfrom the group consisting of —H, alkyl, alkenyl, alkynyl, aryl,alicyclic, heteroalicyclic, aralkyl, aryloxyalkyl, alkoxyalkyl, —C(O)R³,—S(O)₂R³, —C(O)—OR³, —CONHR³, —NR² ₂, and —OR³, all except H areoptionally substituted; R¹ is independently selected from the groupconsisting of —H, alkyl, aryl, heteroalicyclic where the cyclic moietycontains a carbonate or thiocarbonate, —C(R²)₂-aryl, -alk-aryl,—C(R²)₂OC(O)NR² ₂, —NR²—C(O)—R³, —C(R²)₂—OC(O)R³, —C(R²)₂—O—C(O)OR³,—C(R²)₂OC(O)SR³, -alk-S—C(O)R³, -alk-S—S-alkylhydroxy, and-alk-S—S—S-alkylhydroxy, or together R¹ and R¹ are -alk-S—S-alk- to forma cyclic group, wherein each “alk” is an independently selectedalkylene, or together R¹ and R¹ are

wherein V and W are independently selected from the group consisting ofhydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl,1-alkenyl, 1-alkynyl, and —R⁹; or together V and Z are connected via achain of 3-5 atoms, only one of which can be a heteroatom, to form partof a cyclic group substituted with hydroxy, acyloxy, alkoxycarboxy, oraryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; or together V and W are connected viaa chain of 3 carbon atoms to form part of a cyclic group substitutedwith hydroxy, acyloxy, alkoxycarboxy, alkylthiocarboxy, hydroxymethyl,or aryloxycarboxy attached to a carbon atom that is three atoms from anoxygen attached to the phosphorus; Z is selected from the groupconsisting of —CH₂OH, —CH₂OCOR³, —CH₂OC(O)SR³, —CH₂OCO₂R³, —SR³,—S(O)R³, —CH₂N₃, —CH₂NR² ₂, —CH₂Ar, —CH(Ar)OH, —CH(CH═CR²R²)OH,—CH(C≡CR²)OH, and —R₂; with the provisos that: a) V, Z, W are not all—H; and b) when Z is —R², then at least one of V and W is not —H or —R⁹;R₂ is selected from the group consisting of R³ and —H; R³ is selectedfrom the group consisting of alkyl, aryl, alicyclic, heteroalicyclic,and aralkyl; R⁴ is independently selected from the group consisting of—H, lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,and lower aryl; R⁵ is selected from the group consisting of lower alkyl,lower aryl, lower aralkyl, lower alicyclic, and lower heteroalicyclic;R⁶ is independently selected from the group consisting of —H, and loweralkyl R⁷ is independently selected from the group consisting of —H,lower alkyl, lower alicyclic, lower heteroalicyclic, lower aralkyl,lower aryl, and —C(O)R¹⁰; R⁸ is independently selected from the groupconsisting of —H, lower alkyl, lower aralkyl, lower aryl, loweralicyclic, —C(O)R¹⁰, or together said R¹ groups form a bidendatealkylene; R⁹ is selected from the group consisting of alkyl, aralkyl,alicyclic, and heteroalicyclic; R¹⁰ is selected from the groupconsisting of —H, lower alkyl, —NH₂, lower aryl, and lower perhaloalkyl;R¹¹ is selected from the group consisting of alkyl, aryl, —OH, —NH₂ and—OR³; and pharmaceutically acceptable prodrugs and salts thereof. 38.The method of claim 20 wherein said disease is atherosclerosis. whereinsaid compound is administered orally.
 39. The method of claims 34, 35,20, 38, or 36 wherein said compound is administered orally.